STEM CELLS

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Stem Cell Basi"s

About this document
This primer on stem "ells is intended (or an+one ,ho ,ishes to learn more about the biologi"al properties o( stem "ells- the important #uestions about stem "ells that are the (o"us o( s"ienti(i" resear"h- and the potential use o( stem "ells in resear"h and in treating disease. The primer in"ludes in(ormation about stem "ells deri*ed (rom embr+oni" and non/embr+oni" tissues. Mu"h o( the in(ormation in"luded here is about stem "ells deri*ed (rom human tissues- but some studies o( animal/deri*ed stem "ells are also des"ribed. The 0ational 1nstitutes o( 2ealth (012) de*eloped this primer to help readers understand the ans,ers to #uestions su"h as 3hat are stem "ells4 3hat are the di((erent t+pes o( stem "ells- and ,here do the+ "ome (rom4 3hat is the potential (or ne, medi"al treatments using stem "ells4 3hat resear"h is needed to make su"h treatments a realit+4

I. Introduction: What are stem cells, and why are they important?
Stem cells ha*e the remarkable potential to de*elop into man+ di((erent "ell t+pes in the bod+ during earl+ li(e and gro,th. 1n addition- in man+ tissues the+ ser*e as a sort o( internal repair s+stem- di*iding essentiall+ ,ithout limit to replenish other "ells as long as the person or animal is still ali*e. 3hen a stem "ell di*ides- ea"h ne, "ell has the potential either to remain a stem "ell or be"ome another t+pe o( "ell ,ith a more spe"iali5ed (un"tion- su"h as a mus"le "ell- a red blood "ell- or a brain "ell. Stem "ells are distinguished (rom other "ell t+pes b+ t,o important "hara"teristi"s. 6irst- the+ are unspe"iali5ed "ells "apable o( rene,ing themsel*es through cell divisionsometimes a(ter long periods o( ina"ti*it+. Se"ond- under "ertain ph+siologi" or e7perimental "onditions- the+ "an be indu"ed to be"ome tissue/ or organ/spe"i(i" "ells

,ith spe"ial (un"tions. 1n some organs- su"h as the gut and bone marro,- stem "ells regularl+ di*ide to repair and repla"e ,orn out or damaged tissues. 1n other organsho,e*er- su"h as the pan"reas and the heart- stem "ells onl+ di*ide under spe"ial "onditions. )ntil re"entl+- s"ientists primaril+ ,orked ,ith t,o kinds o( stem "ells (rom animals and humans embryonic stem cells and non/embr+oni" somatic or adult stem cells. The (un"tions and "hara"teristi"s o( these "ells ,ill be e7plained in this do"ument. S"ientists dis"o*ered ,a+s to deri*e embr+oni" stem "ells (rom earl+ mouse embr+os nearl+ 80 +ears ago- in 19$1. The detailed stud+ o( the biolog+ o( mouse stem "ells led to the dis"o*er+in 199$- o( a method to deri*e stem "ells (rom human embr+os and gro, the "ells in the laborator+. These "ells are "alled human embryonic stem cells. The embr+os used in these studies ,ere "reated (or reprodu"ti*e purposes through in vitro fertili ation pro"edures. 3hen the+ ,ere no longer needed (or that purpose- the+ ,ere donated (or resear"h ,ith the in(ormed "onsent o( the donor. 1n 200:- resear"hers made another breakthrough b+ identi(+ing "onditions that ,ould allo, some spe"iali5ed adult "ells to be ;reprogrammed< geneti"all+ to assume a stem "ell/like state. This ne, t+pe o( stem "ell- "alled induced pluripotent stem cells !I"S#s$- ,ill be dis"ussed in a later se"tion o( this do"ument. Stem "ells are important (or li*ing organisms (or man+ reasons. 1n the 8/ to &/da+/old embr+o- "alled a blastocyst- the inner "ells gi*e rise to the entire bod+ o( the organismin"luding all o( the man+ spe"iali5ed "ell t+pes and organs su"h as the heart- lung- skinsperm- eggs and other tissues. 1n some adult tissues- su"h as bone marro,- mus"le- and brain- dis"rete populations o( adult stem "ells generate repla"ements (or "ells that are lost through normal ,ear and tear- in=ur+- or disease. >i*en their uni#ue regenerati*e abilities- stem "ells o((er ne, potentials (or treating diseases su"h as diabetes and heart disease. 2o,e*er- mu"h ,ork remains to be done in the laborator+ and the "lini" to understand ho, to use these "ells (or cell%based therapies to treat disease- ,hi"h is also re(erred to as re&enerative or reparative medicine. Laborator+ studies o( stem "ells enable s"ientists to learn about the "ells? essential properties and ,hat makes them di((erent (rom spe"iali5ed "ell t+pes. S"ientists are alread+ using stem "ells in the laborator+ to s"reen ne, drugs and to de*elop model s+stems to stud+ normal gro,th and identi(+ the "auses o( birth de(e"ts. @esear"h on stem cells "ontinues to ad*an"e kno,ledge about ho, an organism de*elops (rom a single "ell and ho, health+ "ells repla"e damaged "ells in adult organisms. Stem "ell resear"h is one o( the most (as"inating areas o( "ontemporar+ biolog+- but- as ,ith man+ e7panding (ields o( s"ienti(i" in#uir+- resear"h on stem "ells raises s"ienti(i" #uestions as rapidl+ as it generates ne, dis"o*eries.

II. What are the uni'ue properties of all stem cells?

Stem cells di((er (rom other t+pes o( "ells in the bod+. !ll stem "ellsAregardless o( their sour"eAha*e three general properties 1) the+ are "apable o( di*iding and rene,ing themsel*es (or long periodsB 2) the+ are unspe"iali5edB and 8) the+ "an gi*e rise to spe"iali5ed "ell t+pes. Stem cells are capable of dividing and renewing themselves for long periods. )nlike mus"le "ells- blood "ells- or ner*e "ellsA,hi"h do not normall+ repli"ate themsel*esAstem "ells ma+ repli"ate man+ times- or proliferate. ! starting population o( stem "ells that proli(erates (or man+ months in the laborator+ "an +ield millions o( "ells. 1( the resulting "ells "ontinue to be unspe"iali5ed- like the parent stem "ells- the "ells are said to be "apable o( lon&%term self%renewal. S"ientists are tr+ing to understand t,o (undamental properties o( stem "ells that relate to their long/term sel(/rene,al 1. 3h+ "an embryonic stem cells proli(erate (or a +ear or more in the laborator+ ,ithout di((erentiating- but most non/embr+oni" stem "ells (adult stem cells) "annotB and 2. 3hat (a"tors in li*ing organisms normall+ regulate stem "ell proli(eration and sel(/rene,al4 Cis"o*ering the ans,ers to these #uestions ma+ make it possible to understand ho, "ell proli(eration is regulated during normal embr+oni" de*elopment or during the abnormal cell division that leads to "an"er. Su"h in(ormation ,ould also enable s"ientists to gro, embr+oni" and non/embr+oni" stem "ells more e((i"ientl+ in the laborator+. The spe"i(i" (a"tors and "onditions that allo, stem "ells to remain unspe"iali5ed are o( great interest to s"ientists. 1t has taken man+ +ears o( trial and error to learn to deri*e and maintain stem "ells in the laborator+ ,ithout them spontaneousl+ di((erentiating into spe"i(i" "ell t+pes. 6or e7ample- it took t,o de"ades to learn ho, to gro, human embryonic stem cells in the laborator+ (ollo,ing the de*elopment o( "onditions (or gro,ing mouse stem "ells. There(ore- understanding the signals in a mature organism that "ause a stem "ell population to proli(erate and remain unspe"iali5ed until the "ells are needed. Su"h in(ormation is "riti"al (or s"ientists to be able to gro, large numbers o( unspe"iali5ed stem "ells in the laborator+ (or (urther e7perimentation. Stem cells are unspecialized. One o( the (undamental properties o( a stem "ell is that it does not ha*e an+ tissue/spe"i(i" stru"tures that allo, it to per(orm spe"iali5ed (un"tions. 6or e7ample- a stem "ell "annot ,ork ,ith its neighbors to pump blood through the bod+

(like a heart mus"le "ell)- and it "annot "arr+ o7+gen mole"ules through the bloodstream (like a red blood "ell). 2o,e*er- unspe"iali5ed stem "ells "an gi*e rise to spe"iali5ed "ellsin"luding heart mus"le "ells- blood "ells- or ner*e "ells. Stem cells can give rise to specialized cells. 3hen unspe"iali5ed stem "ells gi*e rise to spe"iali5ed "ells- the pro"ess is "alled differentiation. 3hile di((erentiating- the "ell usuall+ goes through se*eral stages- be"oming more spe"iali5ed at ea"h step. S"ientists are =ust beginning to understand the signals inside and outside "ells that trigger ea"h step o( the di((erentiation pro"ess. The internal si&nals are "ontrolled b+ a "ell?s &enes- ,hi"h are interspersed a"ross long strands o( C0!- and "arr+ "oded instru"tions (or all "ellular stru"tures and (un"tions. The e7ternal signals (or "ell di((erentiation in"lude "hemi"als se"reted b+ other "ells- ph+si"al "onta"t ,ith neighboring "ells- and "ertain mole"ules in the microenvironment. The intera"tion o( signals during di((erentiation "auses the "ell?s C0! to a"#uire epi&enetic marks that restri"t C0! e7pression in the "ell and "an be passed on through "ell di*ision. Man+ #uestions about stem "ell di((erentiation remain. 6or e7ample- are the internal and e7ternal signals (or "ell di((erentiation similar (or all kinds o( stem "ells4 Can spe"i(i" sets o( signals be identi(ied that promote di((erentiation into spe"i(i" "ell t+pes4 !ddressing these #uestions ma+ lead s"ientists to (ind ne, ,a+s to "ontrol stem "ell di((erentiation in the laborator+- thereb+ gro,ing "ells or tissues that "an be used (or spe"i(i" purposes su"h as cell%based therapies or drug s"reening. !dult stem "ells t+pi"all+ generate the "ell t+pes o( the tissue in ,hi"h the+ reside. 6or e7ample- a blood/(orming adult stem "ell in the bone marro, normall+ gi*es rise to the man+ t+pes o( blood "ells. 1t is generall+ a""epted that a blood/(orming "ell in the bone marro,A,hi"h is "alled a hematopoietic stem cellA"annot gi*e rise to the "ells o( a *er+ di((erent tissue- su"h as ner*e "ells in the brain. E7periments o*er the last se*eral +ears ha*e purported to sho, that stem "ells (rom one tissue ma+ gi*e rise to "ell t+pes o( a "ompletel+ di((erent tissue. This remains an area o( great debate ,ithin the resear"h "ommunit+. This "ontro*ers+ demonstrates the "hallenges o( stud+ing adult stem "ells and suggests that additional resear"h using adult stem "ells is ne"essar+ to understand their (ull potential as (uture therapies.

III. What are embryonic stem cells?

A. What stages of early embryonic development are important for generating embryonic stem cells? (mbryonic stem cells- as their name suggests- are deri*ed (rom embr+os. Most embr+oni" stem "ells are deri*ed (rom embr+os that de*elop (rom eggs that ha*e been (ertili5ed in vitroAin an in vitro fertili ation "lini"Aand then donated (or resear"h purposes ,ith in(ormed "onsent o( the donors. The+ are not deri*ed (rom eggs (ertili5ed in a ,oman?s bod+.

B. How are embryonic stem cells grown in the laboratory? >ro,ing "ells in the laborator+ is kno,n as cell culture. 2uman embr+oni" stem "ells (hESCs) are generated b+ trans(erring "ells (rom a preimplantation/stage embr+o into a plasti" laborator+ "ulture dish that "ontains a nutrient broth kno,n as culture medium. The "ells di*ide and spread o*er the sur(a"e o( the dish. The inner sur(a"e o( the "ulture dish is t+pi"all+ "oated ,ith mouse embr+oni" skin "ells that ha*e been treated so the+ ,ill not di*ide. This "oating la+er o( "ells is "alled a feeder layer. The mouse "ells in the bottom o( the "ulture dish pro*ide the "ells a sti"k+ sur(a"e to ,hi"h the+ "an atta"h. !lsothe (eeder "ells release nutrients into the "ulture medium. @esear"hers ha*e de*ised ,a+s to gro, embr+oni" stem "ells ,ithout mouse (eeder "ells. This is a signi(i"ant s"ienti(i" ad*an"e be"ause o( the risk that *iruses or other ma"romole"ules in the mouse "ells ma+ be transmitted to the human "ells. The pro"ess o( generating an embr+oni" stem "ell line is some,hat ine((i"ient- so lines are not produ"ed ea"h time "ells (rom the preimplantation/stage embr+o are pla"ed into a "ulture dish. 2o,e*er- i( the plated "ells sur*i*e- di*ide- and multipl+ enough to "ro,d the dish- the+ are remo*ed gentl+ and plated into se*eral (resh "ulture dishes. The pro"ess o( re/plating or subculturin& the "ells is repeated man+ times and (or man+ months. Ea"h "+"le o( sub"ulturing the "ells is re(erred to as a passa&e. On"e the "ell line is establishedthe original "ells +ield millions o( embr+oni" stem "ells. Embr+oni" stem "ells that ha*e proli(erated in "ell "ulture (or si7 or more months ,ithout di((erentiating- are pluripotentand appear geneti"all+ normal are re(erred to as an embryonic stem cell line. !t an+ stage in the pro"ess- bat"hes o( "ells "an be (ro5en and shipped to other laboratories (or (urther "ulture and e7perimentation.

C. What laboratory tests are used to identify embryonic stem cells? !t *arious points during the pro"ess o( generating embr+oni" stem "ell lines- s"ientists test the "ells to see ,hether the+ e7hibit the (undamental properties that make them embr+oni" stem "ells. This pro"ess is "alled "hara"teri5ation. S"ientists ,ho stud+ human embr+oni" stem "ells ha*e not +et agreed on a standard batter+ o( tests that measure the "ells? (undamental properties. 2o,e*er- laboratories that gro, human embr+oni" stem "ell lines use se*eral kinds o( tests- in"luding >ro,ing and sub"ulturing the stem "ells (or man+ months. This ensures that the "ells are "apable o( long/term gro,th and sel(/rene,al. S"ientists inspe"t the "ultures through a mi"ros"ope to see that the "ells look health+ and remain undifferentiated. )sing spe"i(i" te"hni#ues to determine the presen"e o( trans"ription (a"tors that are t+pi"all+ produ"ed b+ undi((erentiated "ells. T,o o( the most important trans"ription (a"tors are 0anog and O"t%. Trans"ription (a"tors help turn &enes on and o(( at the right time- ,hi"h is an important part o( the pro"esses o( "ell differentiation and embr+oni" de*elopment. 1n this "ase- both O"t% and 0anog are asso"iated ,ith maintaining the stem "ells in an undi((erentiated state- "apable o( sel(/ rene,al. )sing spe"i(i" te"hni#ues to determine the presen"e o( parti"ular "ell sur(a"e markers that are t+pi"all+ produ"ed b+ undi((erentiated "ells. E7amining the "hromosomes under a mi"ros"ope. This is a method to assess ,hether the "hromosomes are damaged or i( the number o( "hromosomes has "hanged. 1t does not dete"t geneti" mutations in the "ells. Cetermining ,hether the "ells "an be re/gro,n- or sub"ultured- a(ter (ree5ing- tha,ing- and re/plating. Testing ,hether the human embr+oni" stem "ells are pluripotent b+ 1) allo,ing the "ells to di((erentiate spontaneousl+ in "ell "ultureB 2) manipulating the "ells so the+ ,ill di((erentiate to (orm "ells "hara"teristi" o( the three &erm layersB or 8) in=e"ting the "ells into a mouse ,ith a suppressed immune s+stem to test (or the (ormation o( a benign tumor "alled a teratoma. Sin"e the mouse?s immune s+stem is suppressed- the in=e"ted human stem "ells are not re=e"ted b+ the mouse immune s+stem and s"ientists "an obser*e gro,th and di((erentiation o( the human stem "ells. Teratomas t+pi"all+ "ontain a mi7ture o( man+ di((erentiated or partl+ di((erentiated "ell t+pesAan indi"ation that the embr+oni" stem "ells are "apable o( di((erentiating into multiple "ell t+pes.

D. How are embryonic stem cells stimulated to differentiate? !s long as the embr+oni" stem "ells in "ulture are gro,n under appropriate "onditions- the+ "an remain undi((erentiated (unspe"iali5ed). But i( "ells are allo,ed to "lump together to (orm embryoid bodies- the+ begin to di((erentiate spontaneousl+. The+ "an (orm mus"le "ells- ner*e "ells- and man+ other "ell t+pes. !lthough spontaneous di((erentiation is a good indi"ator that a "ulture o( embr+oni" stem "ells is health+- the pro"ess is un"ontrolled and there(ore an ine((i"ient strateg+ to produ"e "ultures o( spe"i(i" "ell t+pes. So- to generate "ultures o( spe"i(i" t+pes o( di((erentiated "ellsAheart mus"le "ells- blood "ells- or ner*e "ells- (or e7ampleAs"ientists tr+ to "ontrol the di((erentiation o( embr+oni" stem "ells. The+ "hange the "hemi"al "omposition o( the "ulture medium- alter the sur(a"e o( the "ulture dish- or modi(+ the "ells b+ inserting spe"i(i" genes. Through +ears o( e7perimentation- s"ientists ha*e established some basi" proto"ols or ;re"ipes< (or the directed differentiation o( embr+oni" stem "ells into some spe"i(i" "ell t+pes (6igure 1). (6or additional e7amples o( dire"ted di((erentiation o( embr+oni" stem "ells- re(er to the 012 stem "ell reports a*ailable at http DDstem"ells.nih.go*Din(oD200:reportD and http DD stem"ells.nih.go*Din(oD2001reportD2001report.htm)

2 ! "herese #inslow $igure 1. %irected differentiation of mouse embr&onic stem cells.

'

1( s"ientists "an reliabl+ dire"t the di((erentiation o( embr+oni" stem "ells into spe"i(i" "ell t+pes- the+ ma+ be able to use the resulting- di((erentiated "ells to treat "ertain diseases in the (uture. Ciseases that might be treated b+ transplanting "ells generated (rom human embr+oni" stem "ells in"lude diabetes- traumati" spinal "ord in=ur+- Cu"henne?s mus"ular d+stroph+- heart disease- and *ision and hearing loss.

I). What are adult stem cells?

!n adult stem cell is thought to be an undifferentiated "ell- (ound among di((erentiated "ells in a tissue or organ that "an rene, itsel( and "an di((erentiate to +ield some or all o( the ma=or spe"iali5ed "ell t+pes o( the tissue or organ. The primar+ roles o( adult stem "ells in a li*ing organism are to maintain and repair the tissue in ,hi"h the+ are (ound. S"ientists also use the term somatic stem cell instead o( adult stem "ell- ,here somati" re(ers to "ells o( the bod+ (not the germ "ells- sperm or eggs). )nlike embryonic stem cells- ,hi"h are de(ined b+ their origin ("ells (rom the preimplantation/stage embr+o)- the origin o( adult stem "ells in some mature tissues is still under in*estigation. @esear"h on adult stem "ells has generated a great deal o( e7"itement. S"ientists ha*e (ound adult stem "ells in man+ more tissues than the+ on"e thought possible. This (inding has led resear"hers and "lini"ians to ask ,hether adult stem "ells "ould be used (or transplants. 1n (a"t- adult hematopoieti"- or blood/(orming- stem "ells (rom bone marro, ha*e been used in transplants (or %0 +ears. S"ientists no, ha*e e*iden"e that stem "ells e7ist in the brain and the heart. 1( the di((erentiation o( adult stem "ells "an be "ontrolled in the laborator+- these "ells ma+ be"ome the basis o( transplantation/based therapies. The histor+ o( resear"h on adult stem "ells began about &0 +ears ago. 1n the 19&0sresear"hers dis"o*ered that the bone marro, "ontains at least t,o kinds o( stem "ells. One population- "alled hematopoietic stem cells- (orms all the t+pes o( blood "ells in the bod+. ! se"ond population- "alled bone marrow stromal stem cells (also "alled mesenchymal stem cells- or skeletal stem "ells b+ some) ,ere dis"o*ered a (e, +ears later. These non/ hematopoieti" stem "ells make up a small proportion o( the stromal cell population in the bone marro,- and "an generate bone- "artilage- (at- "ells that support the (ormation o( blood- and (ibrous "onne"ti*e tissue. 1n the 19:0s- s"ientists ,ho ,ere stud+ing rats dis"o*ered t,o regions o( the brain that "ontained di*iding "ells that ultimatel+ be"ome ner*e "ells. Cespite these reports- most s"ientists belie*ed that the adult brain "ould not generate ne, ner*e "ells. 1t ,as not until the 1990s that s"ientists agreed that the adult brain does "ontain stem "ells that are able to generate the brain?s three ma=or "ell t+pesAastrocytes and oli&odendrocytes- ,hi"h are non/neuronal "ells- and neurons- or ner*e "ells.

A. Where are adult stem cells found, and what do they normally do? !dult stem "ells ha*e been identi(ied in man+ organs and tissues- in"luding brain- bone marro,- peripheral blood- blood *essels- skeletal mus"le- skin- teeth- heart- gut- li*ero*arian epithelium- and testis. The+ are thought to reside in a spe"i(i" area o( ea"h tissue ("alled a ;stem "ell ni"he<). 1n man+ tissues- "urrent e*iden"e suggests that some t+pes o( stem "ells are peri"+tes- "ells that "ompose the outermost la+er o( small blood *essels. Stem "ells ma+ remain #uies"ent (non/di*iding) (or long periods o( time until the+ are a"ti*ated b+ a normal need (or more "ells to maintain tissues- or b+ disease or tissue in=ur+. T+pi"all+- there is a *er+ small number o( stem "ells in ea"h tissue- and on"e remo*ed (rom the bod+- their "apa"it+ to di*ide is limited- making generation o( large #uantities o( stem "ells di((i"ult. S"ientists in man+ laboratories are tr+ing to (ind better ,a+s to gro, large #uantities o( adult stem "ells in cell culture and to manipulate them to generate spe"i(i" "ell t+pes so the+ "an be used to treat in=ur+ or disease. Some e7amples o( potential treatments in"lude regenerating bone using "ells deri*ed (rom bone marro, stromade*eloping insulin/produ"ing "ells (or t+pe 1 diabetes- and repairing damaged heart mus"le (ollo,ing a heart atta"k ,ith "ardia" mus"le "ells.

B. What tests are used to identify adult stem cells? S"ientists o(ten use one or more o( the (ollo,ing methods to identi(+ adult stem "ells (1) label the "ells in a li*ing tissue ,ith mole"ular markers and then determine the spe"iali5ed "ell t+pes the+ generateB (2) remo*e the "ells (rom a li*ing animal- label them in "ell "ulture- and transplant them ba"k into another animal to determine ,hether the "ells repla"e (or ;repopulate<) their tissue o( origin. 1mportantl+- it must be demonstrated that a single adult stem "ell "an generate a line o( geneti"all+ identi"al "ells that then gi*es rise to all the appropriate di((erentiated "ell t+pes o( the tissue. To "on(irm e7perimentall+ that a putati*e adult stem "ell is indeed a stem "ell- s"ientists tend to sho, either that the "ell "an gi*e rise to these geneti"all+ identi"al "ells in "ulture- andDor that a puri(ied population o( these "andidate stem "ells "an repopulate or re(orm the tissue a(ter transplant into an animal.

C. What is nown about adult stem cell differentiation? !s indi"ated abo*e- s"ientists ha*e reported that adult stem "ells o""ur in man+ tissues and that the+ enter normal differentiation path,a+s to (orm the spe"iali5ed "ell t+pes o( the tissue in ,hi"h the+ reside. )ormal differentiation pathwa&s of adult stem cells. 1n a li*ing animal- adult stem "ells are a*ailable to di*ide (or a long period- ,hen needed- and "an gi*e rise to mature "ell t+pes that ha*e "hara"teristi" shapes and spe"iali5ed stru"tures and (un"tions o( a parti"ular tissue. The (ollo,ing are e7amples o( di((erentiation path,a+s o( adult stem "ells (6igure 2) that ha*e been demonstrated in vitro or in vivo.

$igure 2. *ematopoietic and stromal stem cell differentiation.

2ematopoieti" stem "ells gi*e rise to all the t+pes o( blood "ells red blood "ells- B l+mpho"+tes- T l+mpho"+tes- natural killer "ellsneutrophils- basophils- eosinophils- mono"+tes- and ma"rophages. *esenchymal stem cells ha*e been reported to be present in man+ tissues. Those (rom bone marro, (bone marro, stromal stem "ellsskeletal stem "ells) gi*e rise to a *ariet+ o( "ell t+pes bone "ells (osteoblasts and osteo"+tes)- "artilage "ells ("hondro"+tes)- (at "ells (adipo"+tes)- and stromal "ells that support blood (ormation. 2o,e*erit is not +et "lear ho, similar or dissimilar mesen"h+mal "ells deri*ed

2 ! "herese #inslow

(rom non/bone marro, sour"es are to those (rom bone marro, stroma. +eural stem cells in the brain gi*e rise to its three ma=or "ell t+pes ner*e "ells (neurons) and t,o "ategories o( non/neuronal "ellsAastrocytes and oli&odendrocytes. Epithelial stem "ells in the lining o( the digesti*e tra"t o""ur in deep "r+pts and gi*e rise to se*eral "ell t+pes absorpti*e "ells- goblet "ells'aneth "ells- and enteroendo"rine "ells. Skin stem "ells o""ur in the basal la+er o( the epidermis and at the base o( hair (olli"les. The epidermal stem "ells gi*e rise to keratino"+tes,hi"h migrate to the sur(a"e o( the skin and (orm a prote"ti*e la+er. The (olli"ular stem "ells "an gi*e rise to both the hair (olli"le and to the epidermis. "ransdifferentiation. ! number o( e7periments ha*e reported that "ertain adult stem "ell t+pes "an di((erentiate into "ell t+pes seen in organs or tissues other than those e7pe"ted (rom the "ells? predi"ted lineage (i.e.- brain stem "ells that di((erentiate into blood "ells or blood/(orming "ells that di((erentiate into "ardia" mus"le "ells- and so (orth). This reported phenomenon is "alled transdifferentiation. !lthough isolated instan"es o( transdi((erentiation ha*e been obser*ed in some *ertebrate spe"ies- ,hether this phenomenon a"tuall+ o""urs in humans is under debate b+ the s"ienti(i" "ommunit+. 1nstead o( transdi((erentiation- the obser*ed instan"es ma+ in*ol*e (usion o( a donor "ell ,ith a re"ipient "ell. !nother possibilit+ is that transplanted stem "ells are se"reting (a"tors that en"ourage the re"ipient?s o,n stem "ells to begin the repair pro"ess. E*en ,hen transdi((erentiation has been dete"ted- onl+ a *er+ small per"entage o( "ells undergo the pro"ess. 1n a *ariation o( transdi((erentiation e7periments- s"ientists ha*e re"entl+ demonstrated that "ertain adult "ell t+pes "an be ;reprogrammed< into other "ell t+pes in vivo using a ,ell/"ontrolled pro"ess o( geneti" modi(i"ation (see Se"tion E1 (or a dis"ussion o( the prin"iples o( reprogramming). This strateg+ ma+ o((er a ,a+ to reprogram a*ailable "ells into other "ell t+pes that ha*e been lost or damaged due to disease. 6or e7ample- one re"ent e7periment sho,s ho, pan"reati" beta "ells- the insulin/produ"ing "ells that are lost or damaged in diabetes- "ould possibl+ be "reated b+ reprogramming other pan"reati" "ells. B+ ;re/starting< e7pression o( three "riti"al beta/"ell genes in di((erentiated adult pan"reati" e7o"rine "ells- resear"hers ,ere able to "reate beta "ellFlike "ells that "an se"rete insulin. The reprogrammed "ells ,ere similar to beta "ells in appearan"e- si5e- and shapeB e7pressed genes "hara"teristi" o( beta "ellsB and ,ere able to partiall+ restore blood sugar regulation in mi"e ,hose o,n beta "ells had been "hemi"all+ destro+ed. 3hile not transdi((erentiation b+ de(inition- this method (or reprogramming adult "ells ma+ be used as a model (or dire"tl+ reprogramming other adult "ell t+pes.

11

1n addition to reprogramming "ells to be"ome a spe"i(i" "ell t+pe- it is no, possible to reprogram adult somati" "ells to be"ome like embr+oni" stem "ells (induced pluripotent stem cells, i"S#s) through the introdu"tion o( embr+oni" genes. Thus- a sour"e o( "ells "an be generated that are spe"i(i" to the donor- thereb+ a*oiding issues o( histo"ompatibilit+- i( su"h "ells ,ere to be used (or tissue regeneration. 2o,e*er- like embr+oni" stem "ells- determination o( the methods b+ ,hi"h i'SCs "an be "ompletel+ and reprodu"ibl+ "ommitted to appropriate "ell lineages is still under in*estigation.

D. What are the ey !uestions about adult stem cells? Man+ important #uestions about adult stem "ells remain to be ans,ered. The+ in"lude 2o, man+ kinds o( adult stem "ells e7ist- and in ,hi"h tissues do the+ e7ist4 2o, do adult stem "ells e*ol*e during de*elopment and ho, are the+ maintained in the adult4 !re the+ ;le(to*er< embr+oni" stem "ells- or do the+ arise in some other ,a+4 3h+ do stem "ells remain in an undi((erentiated state ,hen all the "ells around them ha*e di((erentiated4 3hat are the "hara"teristi"s o( their ;ni"he< that "ontrols their beha*ior4 Co adult stem "ells ha*e the "apa"it+ to transdi((erentiate- and is it possible to "ontrol this pro"ess to impro*e its reliabilit+ and e((i"ien"+4 1( the bene(i"ial e((e"t o( adult stem "ell transplantation is a trophi" e((e"t- ,hat are the me"hanisms4 1s donor "ell/re"ipient "ell "onta"t re#uired- se"retion o( (a"tors b+ the donor "ell- or both4 3hat are the (a"tors that "ontrol adult stem "ell proli(eration and di((erentiation4 3hat are the (a"tors that stimulate stem "ells to relo"ate to sites o( in=ur+ or damage- and ho, "an this pro"ess be enhan"ed (or better healing4

). What are the similarities and differences between embryonic and adult stem cells?
,uman embryonic and adult stem cells ea"h ha*e ad*antages and disad*antages regarding potential use (or cell%based re&enerative therapies. One ma=or di((eren"e bet,een adult and embr+oni" stem "ells is their di((erent abilities in the number and t+pe o( di((erentiated "ell t+pes the+ "an be"ome. (mbryonic stem cells "an be"ome all "ell t+pes o( the bod+ be"ause the+ are pluripotent. !dult stem "ells are thought to be limited to di((erentiating into di((erent "ell t+pes o( their tissue o( origin.

12

Embr+oni" stem "ells "an be gro,n relati*el+ easil+ in "ulture. !dult stem "ells are rare in mature tissues- so isolating these "ells (rom an adult tissue is "hallenging- and methods to e7pand their numbers in cell culture ha*e not +et been ,orked out. This is an important distin"tion- as large numbers o( "ells are needed (or stem "ell repla"ement therapies. S"ientists belie*e that tissues deri*ed (rom embr+oni" and adult stem "ells ma+ di((er in the likelihood o( being re=e"ted a(ter transplantation. 3e don?t +et kno, ,hether tissues deri*ed (rom embr+oni" stem "ells ,ould "ause transplant re=e"tion- sin"e the (irst phase 1 "lini"al trial testing the sa(et+ o( "ells deri*ed (rom hESCS has onl+ re"entl+ been appro*ed b+ the )nited States 6ood and Crug !dministration (6C!). !dult stem "ells- and tissues deri*ed (rom them- are "urrentl+ belie*ed less likel+ to initiate re=e"tion a(ter transplantation. This is be"ause a patient?s o,n "ells "ould be e7panded in "ulture- "oa7ed into assuming a spe"i(i" "ell t+pe (differentiation)- and then reintrodu"ed into the patient. The use o( adult stem "ells and tissues deri*ed (rom the patient?s o,n adult stem "ells ,ould mean that the "ells are less likel+ to be re=e"ted b+ the immune s+stem. This represents a signi(i"ant ad*antage- as immune re=e"tion "an be "ir"um*ented onl+ b+ "ontinuous administration o( immunosuppressi*e drugs- and the drugs themsel*es ma+ "ause deleterious side e((e"ts.

)I. What are induced pluripotent stem cells?

Induced pluripotent stem cells !i"S#s$ are adult "ells that ha*e been geneti"all+ reprogrammed to an embr+oni" stem "ellFlike state b+ being (or"ed to e7press genes and (a"tors important (or maintaining the de(ining properties o( embr+oni" stem "ells. !lthough these "ells meet the de(ining "riteria (or pluripotent stem "ells- it is not kno,n i( i'SCs and embr+oni" stem "ells di((er in "lini"all+ signi(i"ant ,a+s. Mouse i'SCs ,ere (irst reported in 200:- and human i'SCs ,ere (irst reported in late 200G. Mouse i'SCs demonstrate important "hara"teristi"s o( pluripotent stem "ells- in"luding e7pressing stem "ell markers- (orming tumors "ontaining "ells (rom all three germ la+ers- and being able to "ontribute to man+ di((erent tissues ,hen in=e"ted into mouse embr+os at a *er+ earl+ stage in de*elopment. 2uman i'SCs also e7press stem "ell markers and are "apable o( generating "ells "hara"teristi" o( all three &erm layers. !lthough additional resear"h is needed- i'SCs are alread+ use(ul tools (or drug de*elopment and modeling o( diseases- and s"ientists hope to use them in transplantation medi"ine. Eiruses are "urrentl+ used to introdu"e the reprogramming (a"tors into adult "ells- and this pro"ess must be "are(ull+ "ontrolled and tested be(ore the te"hni#ue "an lead to use(ul treatments (or humans. 1n animal studies- the *irus used to introdu"e the stem "ell (a"tors sometimes "auses "an"ers. @esear"hers are "urrentl+ in*estigating non/

13

*iral deli*er+ strategies. 1n an+ "ase- this breakthrough dis"o*er+ has "reated a po,er(ul ne, ,a+ to ;de/di((erentiate< "ells ,hose de*elopmental (ates had been pre*iousl+ assumed to be determined. 1n addition- tissues deri*ed (rom i'SCs ,ill be a nearl+ identi"al mat"h to the "ell donor and thus probabl+ a*oid re=e"tion b+ the immune s+stem. The i'SC strateg+ "reates pluripotent stem "ells that- together ,ith studies o( other t+pes o( pluripotent stem "ells- ,ill help resear"hers learn ho, to reprogram "ells to repair damaged tissues in the human bod+.

)II. What are the potential uses of human stem cells and the obstacles that must be overcome before these potential uses will be reali ed?
There are man+ ,a+s in ,hi"h human stem "ells "an be used in resear"h and the "lini". Studies o( human embryonic stem cells ,ill +ield in(ormation about the "omple7 e*ents that o""ur during human de*elopment. ! primar+ goal o( this ,ork is to identi(+ ho, undifferentiated stem "ells be"ome the di((erentiated "ells that (orm the tissues and organs. S"ientists kno, that turning &enes on and o(( is "entral to this pro"ess. Some o( the most serious medi"al "onditions- su"h as "an"er and birth de(e"ts- are due to abnormal cell division and differentiation. ! more "omplete understanding o( the geneti" and mole"ular "ontrols o( these pro"esses ma+ +ield in(ormation about ho, su"h diseases arise and suggest ne, strategies (or therap+. 'redi"tabl+ "ontrolling "ell proli(eration and di((erentiation re#uires additional basi" resear"h on the mole"ular and geneti" signals that regulate "ell di*ision and spe"iali5ation. 3hile re"ent de*elopments ,ith i'S "ells suggest some o( the spe"i(i" (a"tors that ma+ be in*ol*ed- te"hni#ues must be de*ised to introdu"e these (a"tors sa(el+ into the "ells and "ontrol the pro"esses that are indu"ed b+ these (a"tors. 2uman stem "ells "ould also be used to test ne, drugs. 6or e7ample- ne, medi"ations "ould be tested (or sa(et+ on di((erentiated "ells generated (rom human pluripotent "ell lines. Other kinds o( "ell lines are alread+ used in this ,a+. Can"er "ell lines- (or e7ampleare used to s"reen potential anti/tumor drugs. The a*ailabilit+ o( pluripotent stem "ells ,ould allo, drug testing in a ,ider range o( "ell t+pes. 2o,e*er- to s"reen drugs e((e"ti*el+- the "onditions must be identi"al ,hen "omparing di((erent drugs. There(ores"ientists ,ill ha*e to be able to pre"isel+ "ontrol the di((erentiation o( stem "ells into the spe"i(i" "ell t+pe on ,hi"h drugs ,ill be tested. Current kno,ledge o( the signals "ontrolling di((erentiation (alls short o( being able to mimi" these "onditions pre"isel+ to generate pure populations o( di((erentiated "ells (or ea"h drug being tested. 'erhaps the most important potential appli"ation o( human stem "ells is the generation o( "ells and tissues that "ould be used (or cell%based therapies. Toda+- donated organs and tissues are o(ten used to repla"e ailing or destro+ed tissue- but the need (or transplantable tissues and organs (ar out,eighs the a*ailable suppl+. Stem "ells- dire"ted to di((erentiate

14

into spe"i(i" "ell t+pes- o((er the possibilit+ o( a rene,able sour"e o( repla"ement "ells and tissues to treat diseases in"luding !l5heimer?s disease- spinal "ord in=ur+- stroke- burnsheart disease- diabetes- osteoarthritis- and rheumatoid arthritis. 6or e7ample- it ma+ be"ome possible to generate health+ heart mus"le "ells in the laborator+ and then transplant those "ells into patients ,ith "hroni" heart disease. 'reliminar+ resear"h in mi"e and other animals indi"ates that bone marrow stromal cells- transplanted into a damaged heart- "an ha*e bene(i"ial e((e"ts. 3hether these "ells "an generate heart mus"le "ells or stimulate the gro,th o( ne, blood *essels that repopulate the heart tissue- or help *ia some other me"hanism is a"ti*el+ under in*estigation. 6or e7ample- in=e"ted "ells ma+ repair b+ se"reting gro,th (a"tors- rather than a"tuall+ in"orporating into the heart. 'romising results (rom animal studies ha*e ser*ed as the basis (or a small number o( e7plorator+ studies in humans (see ;Can Stem Cells Mend a Broken 2eart4< on page 1:). Other re"ent studies in cell culture s+stems indi"ate that it ma+ be possible to dire"t the differentiation o( embr+oni" stem "ells or adult bone marro, "ells into heart mus"le "ells (6igure 8).

2 ! "herese #inslow $igure 3. Strategies to repair heart muscle with adult stem cells.

15

#an Stem #ells *end a -ro.en ,eart?: Stem #ells for the /uture 0reatment of ,eart 1isease
Cardio*as"ular disease (CEC)- ,hi"h in"ludes h+pertension- "oronar+ heart diseasestroke- and "ongesti*e heart (ailure- has ranked as the number one "ause o( death in the )nited States e*er+ +ear sin"e 1900 e7"ept 191$- ,hen the nation struggled ,ith an in(luen5a epidemi". 0earl+ 2-:00 !meri"ans die o( CEC ea"h da+- roughl+ one person e*er+ 8% se"onds. >i*en the aging o( the population and the relati*el+ dramati" re"ent in"reases in the pre*alen"e o( "ardio*as"ular risk (a"tors su"h as obesit+ and t+pe 2 diabetes- CEC ,ill be a signi(i"ant health "on"ern ,ell into the 21st "entur+. Cardio*as"ular disease "an depri*e heart tissue o( o7+gen- thereb+ killing "ardia" mus"le "ells ("ardiom+o"+tes). This loss triggers a "as"ade o( detrimental e*entsin"luding (ormation o( s"ar tissue- an o*erload o( blood (lo, and pressure "apa"it+the o*erstret"hing o( *iable "ardia" "ells attempting to sustain "ardia" output- leading to heart (ailure- and e*entual death. @estoring damaged heart mus"le tissue- through repair or regeneration- is there(ore a potentiall+ ne, strateg+ to treat heart (ailure. The use o( embr+oni" and adult/deri*ed stem "ells (or "ardia" repair is an a"ti*e area o( resear"h. ! number o( stem "ell t+pes- in"luding embr+oni" stem (ES) "ells- "ardia" stem "ells that naturall+ reside ,ithin the heart- m+oblasts (mus"le stem "ells)- adult bone marro,Fderi*ed "ells in"luding mesen"h+mal "ells (bone marro,Fderi*ed "ells that gi*e rise to tissues su"h as mus"le- bone- tendons- ligaments- and adipose tissue)endothelial progenitor "ells ("ells that gi*e rise to the endothelium- the interior lining o( blood *essels)- and umbili"al cord blood cells- ha*e been in*estigated as possible sour"es (or regenerating damaged heart tissue. !ll ha*e been e7plored in mouse or rat models- and some ha*e been tested in larger animal models- su"h as pigs. ! (e, small studies ha*e also been "arried out in humans- usuall+ in patients ,ho are undergoing open/heart surger+. Se*eral o( these ha*e demonstrated that stem "ells that are in=e"ted into the "ir"ulation or dire"tl+ into the in=ured heart tissue appear to impro*e "ardia" (un"tion andDor indu"e the (ormation o( ne, "apillaries. The me"hanism (or this repair remains "ontro*ersial- and the stem "ells likel+ regenerate heart tissue through se*eral path,a+s. 2o,e*er- the stem "ell populations that ha*e been tested in these e7periments *ar+ ,idel+- as do the "onditions o( their puri(i"ation and appli"ation. !lthough mu"h more resear"h is needed to assess the sa(et+ and impro*e the e((i"a"+ o( this approa"h- these preliminar+ "lini"al e7periments sho, ho, stem "ells ma+ one da+ be used to repair damaged heart tissue- thereb+ redu"ing the burden o( "ardio*as"ular disease.

16

1n people ,ho su((er (rom t+pe 1 diabetes- the "ells o( the pan"reas that normall+ produ"e insulin are destro+ed b+ the patient?s o,n immune s+stem. 0e, studies indi"ate that it ma+ be possible to dire"t the di((erentiation o( human embr+oni" stem "ells in "ell "ulture to (orm insulin/produ"ing "ells that e*entuall+ "ould be used in transplantation therap+ (or persons ,ith diabetes. To reali5e the promise o( no*el "ell/based therapies (or su"h per*asi*e and debilitating diseases- s"ientists must be able to manipulate stem "ells so that the+ possess the ne"essar+ "hara"teristi"s (or su""ess(ul di((erentiation- transplantation- and engra(tment. The (ollo,ing is a list o( steps in su""ess(ul "ell/based treatments that s"ientists ,ill ha*e to learn to "ontrol to bring su"h treatments to the "lini". To be use(ul (or transplant purposes- stem "ells must be reprodu"ibl+ made to 'roli(erate e7tensi*el+ and generate su((i"ient #uantities o( "ells (or making tissue. Ci((erentiate into the desired "ell t+pe(s). Sur*i*e in the re"ipient a(ter transplant. 1ntegrate into the surrounding tissue a(ter transplant. 6un"tion appropriatel+ (or the duration o( the re"ipient?s li(e. !*oid harming the re"ipient in an+ ,a+. !lso- to a*oid the problem o( immune re=e"tion- s"ientists are e7perimenting ,ith di((erent resear"h strategies to generate tissues that ,ill not be re=e"ted. To summari5e- stem "ells o((er e7"iting promise (or (uture therapies- but signi(i"ant te"hni"al hurdles remain that ,ill onl+ be o*er"ome through +ears o( intensi*e resear"h.

1'

)III. Where can I &et more information?

6or a more detailed dis"ussion o( stem "ells- see the 012?s Stem Cell @eports. Che"k the 6re#uentl+ !sked Huestions page (or #ui"k ans,ers to spe"i(i" #ueries. The (ollo,ing ,ebsites- ,hi"h are not part o( the 012 Stem Cell 1n(ormation site- also "ontain in(ormation about stem "ells. The 012 is not responsible (or the "ontent o( these sites. http DD,,,.iss"r.orgDpubli" Stem "ell in(ormation (or the publi" (rom the 1nternational So"iet+ (or Stem Cell @esear"h (1SSC@). http DD,,,.nlm.nih.go*DmedlineplusDstem"ells.html Medline 'lus is a "onsumer health database that in"ludes ne,s- health resour"es- "lini"al trials- and more. http DD,,,.e7plorestem"ells."o.uk ! )nited IingdomFbased resour"e (or the general publi" that dis"usses the use o( stem "ells in medi"al treatments and therapies. http DD,,,.stem"ellresear"hne,s."om ! "ommer"ial- online ne,sletter that (eatures stories about stem "ells o( all t+pes.

1!

2lossary

#ell divisionAMethod b+ ,hi"h a single "ell di*ides to "reate t,o "ells. There are t,o main t+pes o( "ell di*ision Adult stem cellASee Somatic stem cell. depending on ,hat happens to the "hromosomes mitosis and meiosis. AstrocyteA! t+pe o( supporting (glial) "ell (ound in the ner*ous s+stem. #hromosome3! stru"ture "onsisting -lastocoelAThe (luid/(illed "a*it+ inside o( C0! and regulator+ proteins the blasto"+st- an earl+- preimplantation (ound in the nu"leus o( the "ell. The stage o( the de*eloping embr+o. C0! in the nu"leus is usuall+ di*ided up among se*eral "hromosomes. The number o( "hromosomes in the -lastocystA! preimplantation embr+o o( about 1&0 "ells produ"ed b+ "ell di*ision nu"leus *aries depending on the spe"ies (ollo,ing (ertili5ation. The blasto"+st o( the organism. 2umans ha*e %: is a sphere made up o( an outer la+er "hromosomes. o( "ells (the trophoblast)- a (luid/(illed "a*it+ (the blasto"oel)- and a "luster #loneA(*) To generate identi"al "opies o( "ells on the interior (the inner "ell o( a region o( a C0! mole"ule or to mass). generate geneti"all+ identi"al "opies o( a "ell- or organismB (n) The identi"al mole"ule- "ell- or organism that results -one marrow stromal cellsA! population (rom the "loning pro"ess. o( "ells (ound in bone marro, that are di((erent (rom blood "ells. 1. 1n re(eren"e to C0! To "lone a -one marrow stromal stem cells gene- one (inds the region ,here !s.eletal stem cells$A! multipotent the gene resides on the C0! and subset o( bone marro, stromal "ells "opies that se"tion o( the C0! using able to (orm bone- "artilage- stromal laborator+ te"hni#ues. "ells that support blood (ormation- (at2. 1n re(eren"e to "ells gro,n in a tissue and (ibrous tissue. "ulture dish a "lone is a line o( "ells that is geneti"all+ identi"al to the originating "ell. This "loned line is #ell%based therapiesATreatment in ,hi"h produ"ed b+ "ell di*ision (mitosis) stem "ells are indu"ed to di((erentiate into the spe"i(i" "ell t+pe re#uired to o( the original "ell. 8. 1n re(eren"e or organisms Man+ repair damaged or destro+ed "ells or tissues. natural "lones are produ"ed b+ plants and (mostl+ in*ertebrate) animals. The term "lone ma+ also be #ell cultureA>ro,th o( "ells in vitro in an arti(i"ial medium (or e7perimental used to re(er to an animal produ"ed b+ somati" "ell nu"lear trans(er resear"h. (SC0T) or parthenogenesis.

1(

#lonin&ASee +lone.

(mbryoA1n humans- the de*eloping organism (rom the time o( (ertili5ation until the end o( the eighth ,eek o( #ord blood stem cellsASee ,mbilical cord blood stem cells. gestation- ,hen it is "alled a (etus. #ulture mediumAThe li#uid that "o*ers (mbryoid bodiesA@ounded "olle"tions "ells in a "ulture dish and "ontains o( "ells that arise ,hen embr+oni" nutrients to nourish and support the stem "ells are "ultured in suspension. Embr+oid bodies "ontain "ell t+pes "ells. Culture medium ma+ also in"lude gro,th (a"tors added to produ"e desired deri*ed (rom all 8 germ la+ers. "hanges in the "ells. (mbryonic &erm cellsA'luripotent stem 1ifferentiationAThe pro"ess ,hereb+ "ells that are deri*ed (rom earl+ germ an unspe"iali5ed embr+oni" "ell "ells (those that ,ould be"ome sperm a"#uires the (eatures o( a spe"iali5ed and eggs). Embr+oni" germ "ells (E> "ell su"h as a heart- li*er- or mus"le "ells) are thought to ha*e properties "ell. Ci((erentiation is "ontrolled b+ similar to embr+oni" stem "ells. the intera"tion o( a "ell?s genes ,ith the ph+si"al and "hemi"al "onditions (mbryonic stem cellsA'rimiti*e outside the "ell- usuall+ through (undi((erentiated) "ells deri*ed (rom signaling path,a+s in*ol*ing proteins a &/da+ preimplantation embr+o embedded in the "ell sur(a"e. that are "apable o( di*iding ,ithout di((erentiating (or a prolonged period in "ulture- and are kno,n to de*elop into 1irected differentiationAThe manipulation o( stem "ell "ulture "ells and tissues o( the three primar+ "onditions to indu"e di((erentiation into germ la+ers. a parti"ular "ell t+pe. (mbryonic stem cell lineAEmbr+oni" 1+AACeo7+ribonu"lei" a"id- a "hemi"al stem "ells- ,hi"h ha*e been "ultured (ound primaril+ in the nu"leus o( under in vitro "onditions that allo, "ells. C0! "arries the instru"tions or proli(eration ,ithout di((erentiation (or blueprint (or making all the stru"tures months to +ears. and materials the bod+ needs to (un"tion. C0! "onsists o( both genes and non/ (ndodermAThe innermost la+er o( the gene C0! in bet,een the genes. "ells deri*ed (rom the inner "ell mass o( the blasto"+stB it gi*es rise to lungsother respirator+ stru"tures- and (ctodermAThe outermost germ la+er o( "ells deri*ed (rom the inner "ell mass o( digesti*e organs- or generall+ ;the gut<. the blasto"+stB gi*es rise to the ner*ous s+stem- sensor+ organs- skin- and related (nucleatedA2a*ing had its nu"leus stru"tures. remo*ed.

(pi&eneticA2a*ing to do ,ith the pro"ess b+ ,hi"h regulator+ proteins "an turn genes on or o(( in a ,a+ that "an be passed on during "ell di*ision.

gastrulation- a period ,hen the inner "ell mass be"omes organi5ed into three distin"t "ell la+ers- "alled germ la+ers. The three la+ers are the e"toderm- the mesoderm- and the endoderm.

/eeder layerACells used in "o/"ulture to maintain pluripotent stem "ells. 6or ,ematopoietic stem cellA! stem "ell that human embr+oni" stem "ell "ulturegi*es rise to all red and ,hite blood "ells t+pi"al (eeder la+ers in"lude mouse and platelets. embr+oni" (ibroblasts (ME6s) or human embr+oni" (ibroblasts that ha*e ,uman embryonic stem cell !h(S#$A been treated to pre*ent them (rom ! t+pe o( pluripotent stem "ell deri*ed di*iding. (rom earl+/stage human embr+os- up to and in"luding the blasto"+st stage. hESCs are "apable o( di*iding ,ithout /ertili ationAThe =oining o( the male gamete (sperm) and the (emale gamete di((erentiating (or a prolonged period in (egg). "ulture and are kno,n to de*elop into "ells and tissues o( the three primar+ germ la+ers. See also pluripotent/etusA1n humans- the de*eloping human (rom appro7imatel+ eight ,eeks a(ter blastoc&st- and germ la&ers. "on"eption until the time o( its birth. Induced pluripotent stem cell !i"S#$A 2ameteA!n egg (in the (emale) or sperm ! t+pe o( pluripotent stem "ell- similar (in the male) "ell. See also Somatic cell. to an embr+oni" stem "ell- (ormed b+ the introdu"tion o( "ertain embr+oni" genes into a somati" "ell. 2astrulationAThe pro"ess in ,hi"h "ells proli(erate and migrate ,ithin the embr+o to trans(orm the inner "ell mass "n vitroALatin (or ;in glass<B in a o( the blasto"+st stage into an embr+o laborator+ dish or test tubeB an arti(i"ial "ontaining all three primar+ germ en*ironment. la+ers. "n vitro fertili ationA! te"hni#ue that 2eneA! (un"tional unit o( heredit+ unites the egg and sperm in a laborator+that is a segment o( C0! (ound on instead o( inside the (emale bod+. "hromosomes in the nu"leus o( a "ell. >enes dire"t the (ormation o( an Inner cell mass !I#*$AThe "luster o( "ells en5+me or other protein. inside the blasto"+st. These "ells gi*e rise to the embr+o and ultimatel+ the (etus. The 1CM "ells ma+ be used to generate 2erm layersA!(ter the blasto"+st stage o( embr+oni" de*elopment- the inner embr+oni" stem "ells. "ell mass o( the blasto"+st goes through

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4on&%term self%renewalAThe abilit+ The number o( "hromosomes remains o( stem "ells to rene, themsel*es b+ the same in this t+pe o( "ell di*ision. di*iding into the same non/spe"iali5ed "ell t+pe o*er long periods (man+ *ultipotentA2a*ing the abilit+ to de*elop months to +ears) depending on the into more than one "ell t+pe o( the spe"i(i" t+pe o( stem "ell. bod+. See also pluripotent and totipotent. *esenchymal stem cellsA! term that +eural stem cellA! stem "ell (ound in is "urrentl+ used to de(ine non/blood adult neural tissue that "an gi*e rise to adult stem "ells (rom a *ariet+ o( neurons and glial (supporting) "ells. tissues- although it is not "lear that E7amples o( glial "ells in"lude astro"+tes mesen"h+mal stem "ells (rom di((erent and oligodendro"+tes. tissues are the same. +euronsA0er*e "ells- the prin"ipal *eiosisAThe t+pe o( "ell di*ision a (un"tional units o( the ner*ous s+stem. diploid germ "ell undergoes to produ"e ! neuron "onsists o( a "ell bod+ and gametes (sperm or eggs) that ,ill "arr+ its pro"essesAan a7on and one or hal( the normal "hromosome number. more dendrites. 0eurons transmit This is to ensure that ,hen (ertili5ation in(ormation to other neurons or "ells b+ o""urs- the (ertili5ed egg ,ill "arr+ the releasing neurotransmitters at s+napses. normal number o( "hromosomes rather than "ausing aneuploid+ (an abnormal 5li&odendrocyteA! supporting "ell that number o( "hromosomes). pro*ides insulation to ner*e "ells b+ (orming a m+elin sheath (a (att+ la+er) around a7ons. *esodermAMiddle la+er o( a group o( "ells deri*ed (rom the inner "ell mass o( the blasto"+stB it gi*es rise to bone- "artheno&enesisAThe arti(i"ial a"ti*ation mus"le- "onne"ti*e tissue- kidne+s- and o( an egg in the absen"e o( a spermB the related stru"tures. egg begins to di*ide as i( it has been (ertili5ed. *icroenvironmentAThe mole"ules and "ompounds su"h as nutrients and "assa&eA1n "ell "ulture- the pro"ess in gro,th (a"tors in the (luid surrounding ,hi"h "ells are disasso"iated- ,asheda "ell in an organism or in the and seeded into ne, "ulture *essels laborator+- ,hi"h pla+ an important a(ter a round o( "ell gro,th and role in determining the "hara"teristi"s proli(eration. The number o( passages a o( the "ell. line o( "ultured "ells has gone through is an indi"ation o( its age and e7pe"ted stabilit+. *itosisAThe t+pe o( "ell di*ision that allo,s a population o( "ells to in"rease its numbers or to maintain its numbers.

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"luripotentA2a*ing the abilit+ to gi*e preimplantation stage embr+os (ertili5ed rise to all o( the *arious "ell t+pes o( outside a ,oman?s bod+ .in vitro/. the bod+. 'luripotent "ells "annot make e7tra/embr+oni" tissues su"h "roliferationAE7pansion o( the number as the amnion- "horion- and other o( "ells b+ the "ontinuous di*ision o( "omponents o( the pla"enta. S"ientists single "ells into t,o identi"al daughter demonstrate pluripoten"+ b+ pro*iding "ells. e*iden"e o( stable de*elopmental potential- e*en a(ter prolonged 6e&enerative medicineA! (ield o( "ulture- to (orm deri*ati*es o( all medi"ine de*oted to treatments three embr+oni" germ la+ers (rom the in ,hi"h stem "ells are indu"ed to progen+ o( a single "ell and to generate di((erentiate into the spe"i(i" "ell t+pe a teratoma a(ter in=e"tion into an re#uired to repair damaged or destro+ed immunosuppressed mouse. "ell populations or tissues. See also cell0 based therapies. "olar -odyA! polar bod+ is a stru"ture produ"ed ,hen an earl+ egg "ell- or 6eproductive clonin&AThe pro"ess o( using somati" "ell nu"lear trans(er oogonium- undergoes meiosis. 1n the (irst meiosis- the oogonium di*ides its (SC0T) to produ"e a normal- (ull "hromosomes e*enl+ bet,een the t,o gro,n organism (e.g.- animal) "ells but di*ides its "+toplasm une#uall+. geneti"all+ identi"al to the organism One "ell retains most o( the "+toplasm(animal) that donated the somati" ,hile the other gets almost none"ell nu"leus. 1n mammals- this ,ould lea*ing it *er+ small. This smaller "ell re#uire implanting the resulting embr+o is "alled the (irst polar bod+. The (irst in a uterus ,here it ,ould undergo polar bod+ usuall+ degenerates. The normal de*elopment to be"ome a li*e o*um- or larger "ell- then di*ides againindependent being. The (irst animal produ"ing a se"ond polar bod+ ,ith to be "reated b+ reprodu"ti*e "loning hal( the amount o( "hromosomes but ,as Coll+ the sheep- born at the @oslin almost no "+toplasm. The se"ond polar 1nstitute in S"otland in 199:. See also bod+ splits o(( and remains ad=a"ent Somatic cell nuclear transfer .S+)"/. to the large "ell- or oo"+te- until it (the se"ond polar bod+) degenerates. Onl+ Si&nalsA1nternal and e7ternal (a"tors one large (un"tional oo"+te- or egg- is that "ontrol "hanges in "ell stru"ture produ"ed at the end o( meiosis. and (un"tion. The+ "an be "hemi"al or ph+si"al in nature. "reimplantationA3ith regard to an embr+o- preimplantation means that Somatic cellA!n+ bod+ "ell other than the embr+o has not +et implanted gametes (egg or sperm)B sometimes in the ,all o( the uterus. 2uman re(erred to as ;adult< "ells. See also embr+oni" stem "ells are deri*ed (rom 1amete.

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Somatic cell nuclear transfer !S#+0$A immune s+stem. S"ientists test ,hether ! the+ ha*e established a human te"hni#ue that "ombines an enu"leated embr+oni" stem "ell (hESC) line b+ egg and the nu"leus o( a somati" "ell to in=e"ting putati*e stem "ells into su"h make an embr+o. SC0T "an be used (or mi"e and *eri(+ing that the resulting therapeuti" or reprodu"ti*e purposesteratomas "ontain "ells deri*ed (rom all but the initial stage that "ombines three embr+oni" germ la+ers. an enu"leated egg and a somati" "ell nu"leus is the same. See also "herapeutic 0herapeutic clonin&AThe pro"ess o( cloning and 2eproductive cloning. using somati" "ell nu"lear trans(er (SC0T) to produ"e "ells that e7a"tl+ Somatic !adult$ stem cellA! relati*el+ mat"h a patient. B+ "ombining a rare undi((erentiated "ell (ound in man+ patient?s somati" "ell nu"leus and an organs and di((erentiated tissues ,ith enu"leated egg- a s"ientist ma+ har*est a limited "apa"it+ (or both sel( rene,al embr+oni" stem "ells (rom the resulting (in the laborator+) and di((erentiation. embr+o that "an be used to generate Su"h "ells *ar+ in their di((erentiation tissues that mat"h a patient?s bod+. This "apa"it+- but it is usuall+ limited to "ell means the tissues "reated are unlikel+ t+pes in the organ o( origin. This is an to be re=e"ted b+ the patient?s immune a"ti*e area o( in*estigation. s+stem. See also Somatic cell nuclear transfer .S+)"/. Stem cellsACells ,ith the abilit+ to di*ide (or inde(inite periods in "ulture and to 0otipotentA2a*ing the abilit+ to gi*e gi*e rise to spe"iali5ed "ells. rise to all the "ell t+pes o( the bod+ plus all o( the "ell t+pes that make up Stromal cellsAConne"ti*e tissue "ells (ound the e7traembr+oni" tissues su"h as in *irtuall+ e*er+ organ. 1n bone marro,the pla"enta. See also 3luripotent and stromal "ells support blood (ormation. 4ultipotent. Subculturin&ATrans(erring "ultured "ells,ith or ,ithout dilution- (rom one 0ransdifferentiationAThe pro"ess b+ "ulture *essel to another. ,hi"h stem "ells (rom one tissue di((erentiate into "ells o( another tissue. Surface mar.ersA'roteins on the outside sur(a"e o( a "ell that are uni#ue to 0rophectodermAThe outer la+er o( the "ertain "ell t+pes and that "an be preimplantation embr+o in mi"e. 1t *isuali5ed using antibodies or other "ontains trophoblast "ells. dete"tion methods. 0rophoblastAThe outer "ell la+er o( 0eratomaA! multi/la+ered benign tumor the blasto"+st. 1t is responsible (or that gro,s (rom pluripotent "ells implantation and de*elops into the in=e"ted into mi"e ,ith a d+s(un"tional e7traembr+oni" tissues- in"luding the

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pla"enta- and "ontrols the e7"hange o( o7+gen and metabolites bet,een mother and embr+o. 7mbilical cord blood stem cellsAStem "ells "olle"ted (rom the umbili"al "ord at birth that "an produ"e all o( the blood "ells in the bod+ (hematopoieti"). Cord blood is "urrentl+ used to treat patients ,ho ha*e undergone "hemotherap+ to destro+ their bone marro, due to "an"er or other blood/related disorders. 7ndifferentiatedA! "ell that has not +et de*eloped into a spe"iali5ed "ell t+pe.

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).S. Cepartment o( 2ealth and 2uman Ser*i"es 0ational 1nstitutes o( 2ealth