10.

Matine elije

Uvod
ta su matine elije?
Adultne matine elije
Matine elije raka
Pluripotentne matine elije

Matine elije se definiu po

funkcionalnosti
Obnavljanje

Diferencijacija

(funkcionalne identine kopije)

(specializacija ili sazrevanje)

Totipotente = sve elije u placenti i embrionu

Pluripotentne = sve elije u embrionu
ektoderm, mezoderm, endoderm
Multipotentne = grupa elija u datom tkivu
Unipotentne= jedan tip elija
Nullipotentne = nemaju sposobnost diferenciranja

Adultne

Rani razvoj

B
KRV

WBC

Embrionalne

RBC

Fetus

Plts
MezenhimVezivno

Zigot

Kost

Blastocista

Mii
Neural

Implantacija

Neuroni Mast
Oligoglija
Astroglija
jetra, pankreas, koa
testis, creva

Veliki potencijal

diferencijacija

Veliko ogranienje

Adultne
matine
elije

elije iz kotane sri

Adultne matine elije

Zamenjuju I popravljaju

Koa

Stvaraju elije tkiva u kojem

su naene

Mozak

Krv

Skeletna
Muskulatura

Kornealna epitelna
matina elija

Jetra (?)

Adultne matine elija stvaraju

specifino tkivo
Mozak

Koa

Krv

Skeletna
Muskulatura

Kornealne epitelne
matine elije

Jetra (?)

Adultne matine elija mogu da preu u

matine elije raka
neuroblastom

melanom

leukemia

rabdomiosarkom

retinoblastom

HCC

Bolesti matinih elija

Kotana sr
Fankoni anemia
Aplasticna anemia
Diamond-Blackfan anemia
Geneticka imunodeficijencija

Nedostatak rezervi matinih elija

Tipe 1 diabetes
Jetra
Plua
Srce

Opekotine
Gubitak kose
Neurodegeneracije

Maligniteti
Teratomi
Leukemia
Vilmov tumor

Neuroblastoma
Rhabdomyosarcoma

ta je potrebno da se matine elije primene u praksi ?

Genetski identitet (HLA)

Produkcija u klinikoj koliini
istoa
GMP
Potpuno diferencirane elije
Sigurne po pacijenta

Adekvatan prenos do mesta

Efikasno usaivanje
Problem oiljnog tkiva

Funkcionalnost
Integracija

VREME

Poetak
elijske terapije
1667, Engleska
Prva transfuzija
odprilike
Artur Koga dobio
transfuziju jagnjee
krvi da se izlei od
sifilisa
Pacijent preminuo
neto kasnije.

Pioniri matinih elija

Valentin Haker I njegova ilustracija

matinih elija (Stammzelle or st) 1892
Artur Papenhajm 1905: dijagram
razvoja elija krvi

Nakon 1800

Ramalho-Santos and Willenbring, Cell Stem Cell, 2007

Pioniri
1900, Karl Landsteiner krvne grupe (A, B, O)
1915, Riard Levizon, razvoj antikoagulanta

Dr. Riard Levizon

1930

Razvoj aplikacije krvnih matinih elija

December 2, 1943
Bari Harbor, Italy German air raid
US John Harvey was hit and exploded
many tons of mustard gas-filled munitions
extensive leukopenia
seminal papers in JAMA 1919 and 1946

August 6 and 9, 1945

Hiroshima and Nagasaki
Little Boy = uranium
Fat Man = plutonium
acute radiation syndr. (ARS)
extensive leukopenia

Pronalazak elijskih antigena

1960
Dr. Peter Medavar

Transplantacija

1990
Joseph E. Murray

E. Donnall Thomas

1954 (transplantacija bubrega) and 1957 (kotana sr)

N Engl J Med 257, 491-496. 1957

Autologa i Alogena Transplantacija

Autologa = dobija sopstvene elije

Alogenetska = dobija neije tue

Infuzija kotane sri

Stanley Sclavos (www.sclavosnet.com)

Kliniki podaci
3 godine preivljavanja (Kaplan-Meier) 3% 7% sa
nerodbinskim davaocem**
Imunoloke potekoe
Infekcije, katarakta, disfunkcija tireoideje, osteoporoza

2-godine nerodbinski davaoci elije iz pupane

vrpce 42% 5%**
10-godina autologa trasnplantacija 68.8 % 1.8%**

James Till

1960-63

Ernest McCulloch

Kako dokazati da matine elije postoje?

CFU-S

Neuronalna regeneracija

1980

Transplantacija matinih elija

iz pupane vrpce

Privatne i javne banke matinih elija

Nedovoljan broj elija ako je u pitanju I veliko dete
Multi-cord transplantacija kod odraslih

1989

Neuronala regeneracija ljudskog

CNS-a

1998

Cells in the dentate gyrus labeled in vivo with BrdU

Transplantacija beta elija

Prior efforts showed that <10% of transplanted patients

were insulin injection-free at one year
Seven initial patients
At least 2 donor pancreases (>11,000 islets/kg) (some more)
Range of follow-up was 5 months to 15 months
None had been rejected, none had hypoglycemic episodes

2000

2006

Matine elije raka

Kad dobre elije postanu
loe

Transplantacija elija raka kod ivotinja

1938

Earliest human/animal
experiments a curiosity
In early 20th Century
became a tool to primarily
study cancer
1st experiments into sites
of immune privilege (eye)
Subsequent work with
immune suppressed or
athymic
Give mice exp. Rx, etc

1957

Transplantacija matinih
elija kod dobrovoljaca

If they were not immediately

excised, the implants usually
regressed spontaneously and
completely by 4 to 6 weeks.
However, in four patients there
was recurrence of cancer growth
after biopsy at several implant
sites. Three of these recurrences
were completely removed by a
repeat excision on the 19th,
42nd, and 77th days,
respectively. In two patients,
some of the recurrent nodules
grew progressively until the
patients died

Imunodeficijentni mievi

SCID mievi u kasnim

osamdesetim

Pluripotentne matine elije

stvaraju adultne matine elije
Mozak

Koa

Krv

Skeletna
muskulatura

Kornealne epitelne
matine elije

Jetra (?)

Adultne

Rani razvoj

B
KRV

WBC

Embrionalne

RBC

Fetus

Plts
MezenhimVezivno

Zigot

Kost

Blastocista

Mii
Neural

Implantacija

Neuroni Mast
Oligoglija
Astroglija
jetra, pankreas, koa
testis, creva

Veliki potencijal

diferencijacija

Veliko ogranienje

Nuklearno reprogramiranje i razvoj

1920

Hans Spemann
(1869 1941)

Nuklearna jednakost

Hipoteza:
erke elije se
genetski ne razlikuju

Epigenetski
mehanizam

Specijaliziranost tkiva kroz

epigenezu
Organizacija

Jedinice za izgradnju

Remodeliranje hromatina kroz razvoj

Metilacija DNK i acetilizacija histona
+ methylation - acetylation

Gene je off

Gene je on
- methylation + acetylation
Reik and Walter, Nature Reviews Genetics, Vol 2, Jan 2001, 21-32.

Nuklearni transfer
PNAS

~32%

1952

John Gurdon

at Oldham and District General Hospital

July 25, 1978
5 lb 12 oz

> 4,000,000 IVF babies

worldwide

1978

Bob Edwards
Nobel, 2010

ta je blastocista?
~150 do 200 elija
zona pellucida (membrana)

unutranja elijska masa

(embrio/plod)
Trofoblast (placenta)

Samo dva elijska tipa u blastocisti

Prva derivacija embrionalnih

matinih elija

1981

# oocytes used
# blastocysts

277/29/1
# Dollys

0.36%
Dolly
Prvi sisar dobijen
putem kloniranje

1997

Science, 6 November 1998

5/14
~36%

1998

Michael West, M.D., Ph.D.

GERON

Ljudske embrionalne maticne celije

Reprogramiranje putem nuklearnog

transfera

c.1

c.2

c.3

Prado-Lopez et al., 2010

Set od 24 pluripotentnih gena

4 gena

(Oct4/Sox2/Klf4/cMyc)

elija fibroblasta
induced Pluripotent
Stem Cells (iPSC)

2006

Reprogramiranje

Taken from: Yamanaka, Cell Stem Cell, 2007

Humane iPS celije

Sve u 2007-oj

Kako funkcionie?
Skin biopsy is taken from patient
Skin cells are reprogrammed by
viral transduction of 2-7 key
transcription factors
Cells that look and behave like
embryonic stem cells are obtained

Lako et al. (2010) Stem Cells 28: 845-850

Prednosti iPSC

elije koje su uzete od pacijenta

Stvaranje modela bolesti
Razumevanje patologije bolesti
Regeneracija putem transplantacije
Razvoj lekova

Razlicite linije iPS celija koje nose informacije

razlicitih bolesti
2008
(naslednih 1800)

iPSC imaju iste markere kao i embrionalne

matine elije
(TRA1-60, TRA-80, OCT4, NANOG, SOX2,
REX)

Armstrong et al. Stem Cells. 2010 Apr;28(4):661-73

iPSC i embrionalne elije nisu identine

* Gene expression in iPS cells becomes more ES-like over time

* Differences remain - what will these differences mean for research?

Spermatogeneza i
zdravo potomstvo od
matinih elija

Oogeneza, embrionalni razvoj,

i potomstvo od matinih elija

Alchajmerova bolest i maticne

celije

Kicmeni diskovi

Makularna degeneracija

Misicna distrofija

Parkinsonova bolest

Mozdani udar

Povreda kicmene mozdine

Specificno I nespecificno
dejstvo maticnih celija
Specificno dejstvo: preuzimanje specificne
funkcije degenerisanih ili nefunkcionalnih celija
Nespecificno: sekrecija parakrinih faktora koji
stimulisu rast endogenih maticnih celija

Limbalne maticne celije

Hemijske-termalne
povrede
Radijacija
Imunoloska stanja
Tumori oka
Kontaktna sociva kao
uzrok

Limbus

cornea

conjuctiva
limbus

Ocular surface
epithelium
Transit amplifying cells

Limbal stem cells (p63, vimentin, LNGFR)

Bolesti pluca i bioinjzinjering

1 Trachea is removed from dead donor patient

2 It is flushed with chemicals to remove all existing cells
3 Donor trachea "scaffold" coated with stem cells from the patient's hip bone marrow.
Cells from the airway lining added
4 Once cells have grown (after about four days) donor trachea is inserted into patient's bronchus

Transplantacija traheje

A pre-surgery CT
the arrow showing the windpipe narrowing

The trachea graft ready for transplantation

Obecanja i ozbiljni posao

> 2800 studije koje opisuju

razvoj i klinicke pokusaje
primene maticnih celija

Modani tumori nakon transplantacije

neuronalnih matinih elija kod pacijenta sa
Ataxia Telangiectasia
PLoS Med. 2009 Feb 17;6(2):e1000029.

Pomo pacijentima irom sveta

http://www.closerlookatstemcells.org