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Tissue culture
Tissue is a cellular organizational level intermediate between cells and a complete organism. A tissue is an ensemble of similar cells from the same origin that together carry out a specific function. Organs are then formed by the functional grouping together of multiple tissues. Tissue culture generally refers to the growth of cells from a tissue from a multicellular organism in vitro. These cells may be cells isolated from a donor organism, primary cells, or an immortalised cell line. PLANT TISSUE CULTURE Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. EXPLANT- it is any portion from a plant that will be used to initiate a culture. TYPES OF TISSUE CULTURE ORGAN CULTURE: These are cultures of isolated plant organs including cultures derived from root tips , stem tips , leaf primordia or immature part of flowers and immature fruits. Organ culture is a development from tissue culture methods of research; the organ culture is able to accurately model functions of an organ in various states and conditions by the use of the actual in vitro organ itself. EMBRYO CULTURE:- These are culture of isolated immature or mature embryos. Embryo culture has been used to produce plants from embryos that would not normally develop within the fruit. This occurs in early-ripening peaches and in some hybridization between species. Embryo culture can also be used to circumvent seed dormancy. CALLUS OR TISSUE CULTURE:- These are culture of tissue arising from disorganized proliferation of cells from segment of plant organs . Tissue or callus culture are , generally grown on solid medium as amass of cells. It is the growth of tissue outside an organism in a nutrient medium, or the techniques involved in this process. SUSPENSION CULTURE:- These are often called cell cultures, as they represent a lower level of organization than tissue or callus culture. Suspension culture is in vitro cultures of isolated cells and very small cell groups remaining dispersed as they grow in excited liquid media. Plant cell suspension cultures are widely used in plant biology as a convenient tool for the investigation of a wide range of phenomena, bypassing the structural complexity of the plant organism. SUSPENSION CULTURE:- These are often called cell cultures, as they represent a lower level of organization than tissue or callus culture. Suspension culture is in vitro cultures of isolated cells and very small cell groups remaining dispersed as they grow in excited liquid media.

2 GENETICALLY MODIFIED ORGANISMS GMOs, or genetically modified organisms, are plants or animals that have been genetically engineered with DNA from bacteria, viruses or other plants and animals. These experimental combinations of genes from different species cannot occur in nature or in traditional crossbreeding. Organisms that have been genetically modified include micro-organisms such as bacteria and yeast, insects, plants, fish, and mammals. GMOs are the source of genetically modified foods, and are also widely used in scientific research and to produce goods other than food. The term GMO is very close to the technical legal term, 'living modified organism' defined in the Cartagena Protocol on Biosafety, which regulates international trade in living GMOs (specifically, "any living organism that possesses a novel combination of genetic material obtained through the use of modern biotechnology"). Recombinant DNA technology is a technology that allows DNA to be produced via artificial means. The procedure has been used to change DNA in living organisms and may have even more practical uses in the future. It is an area of medical science that is just beginning to be researched in a concerted effort. Transgenic organisms, a subset of GMOs, are organisms which have inserted DNA that originated in a different species. Genetic is the mutation, insertion, or deletion of genes. When genes are inserted, they usually come from a different species, which is a form of horizontal gene transfer. In nature this can occur when exogenous DNA penetrates the cell membrane for any reason. The general principle of producing a GMO is to alter the genetic material of an organism's genome. This may involve mutating, deleting, or adding genetic material. When genetic material from a different species is added, the resulting DNA is called recombinant DNA and the organism is called a transgenic organism. The first recombinant DNA molecules were produced by Paul Berg in 1972. Organisms that have been altered through genetic engineering techniques are known as genetically modified organisms or GMOs. STEM CELL Stem cell therapy is an intervention strategy that introduces new adult stem cells into damaged tissue in order to treat disease or injury. Many medical researchers believe that stem cell treatments have the potential to change the face of human disease and alleviate suffering. The ability of stem cells to self-renew and give rise to subsequent generations with variable degrees of differentiation capacities, offers significant potential for generation of tissues that can potentially replace diseased and damaged areas in the body, with minimal risk of rejection and side effects.

3 USES Baldness Missing teeth Deafness and vision impairment Amyotrophic lateral sclerosis Graft vs. host disease and Crohn's disease Neural and behavioral birth defects Diabetes Transplantation CANCER TREATMENTS

TYPES OF STEM CELL Induced pluripotent cells (iPS cells) are non-pluripotent cells that were induced to become pluripotent, that is, able to form all cell types of the body. In other words, a cell with a specialized function (for example, a skin cell) was 'reprogrammed' to an unspecialized state similar to that of an embryonic stem cell. Cancer stem cells are defined as a subpopulation of cancer cells that presented a greater renewal potential compared to the other tumor cells and the ability to reform all the characteristics of the parental tumor upon transplantation into immunodeficient mice. Cancer stem cells have been postulated to be responsible for tumor growth and tumor relapse after therapy. Somatic cell nuclear transfer (SCNT) is a technique in which the nucleus of a somatic cell (that is any cell of the body apart from the sperm or egg), is transferred into an egg that has had its original nucleus removed. The egg now has the same DNA, or genetic material, as the donor somatic cell. Given the right signals, the egg can be coaxed into developing as if it had been fertilized. Bioethics is the study of the social, moral and ethical issues in the fields of scientific research, medical treatment and, more generally, in the life sciences. HUMAN ORGAN TRANSPLANT Organ transplantation is the moving of an organ from one body to another (or from a donor site on the patient's own body), for the purpose of replacing the recipient's damaged or failing organ with a working one from the donor site. Organ donors can be living or deceased (previously referred to as cadaveric). Organs that can be transplanted include the heart, kidneys , liver , lungs , pancreas , penis , and intestine . Tissues include bones, tendons, cornea, heart valves, veins, arms, and skin. Worldwide, the kidneys are the most commonly transplanted organs. TYPES OF ORGAN TRANSPLANT 1. Autograft Transplant - this is done with surplus tissue, or tissue that can regenerate, or tissues more desperately needed elsewhere. Sometimes an autograft is done to remove the tissue and then treat it or the person, before returning it >skin grafts, vein extraction >storing blood in advance of surgery. 2. Allograft Transplant - is a transplant of an organ or tissue between two genetically nonidentical members of the same species. Most human tissue and organ transplants are allografts. Due to the genetic difference between the organ and the recipient, the recipient's immune system will identify the organ as foreign and attempt to destroy it, causing transplant rejection . To prevent this, the organ recipient must take immunosuppressant .

4 This dramatically affects the entire immune system, making the body vulnerable to pathogens . 3. Isograft - subset of allografts in which organs or tissues are transplanted from a donor to a genetically identical recipient (such as an identical twin ). Isografts are differentiated from other types of transplants because while they are anatomically identical to allografts, they don't trigger an immune response . 4. Xenograft and xenotransplantation- transplant of organs or tissue from one species to another. An example are porcine heart valve transplants, which are quite common and successful. Another example is attempted piscine-primate (fish to non-human primate) transplant of islet (i.e. pancreatic or insular tissue) tissue. The latter research study was intended to pave the way for potential human use, if successful. However, xenotransplantion is often an extremely dangerous type of transplant because of the increased risk of non-compatibility, rejection, and disease carried in the tissue. 5. Split transplants - sometimes a deceased-donor organ, usually a liver, may be divided between two recipients, especially an adult and a child. This is not usually a preferred option because the transplantation of a whole organ is more successful. 6. Domino transplants - this operation is usually performed on patients with cystic fibrosis because both lungs need to be replaced and it is a technically easier operation to replace the heart and lungs at the same time. As the recipient's native heart is usually healthy, it can be transplanted into someone else needing a heart transplant. That term is also used for a special form of liver transplant in which the recipient suffers from familial amyloidotic polyneuropathy , a disease where the liver slowly produces a protein that damages other organs. This patient's liver can be transplanted into an older patient who is likely to die from other causes before a problem arises. MAJOR ORGANS TRANSPLANTED 1.Thoracic organs organs that are located within or in the thorax. These are: Heart (Deceased-donor only) Lung (Deceased-donor and Living-Donor) Heart/Lung (Deceased-donor and Domino transplant) 2. Abdominal organs - that lies between the thorax and the pelvis and encloses the following: Kidney (Deceased-donor and Living-Donor) Liver (Deceased-donor and Living-Donor) Pancreas (Deceased-donor only) Intestine (Deceased-donor and Living-Donor) Stomach (Deceased-donor only) 3. Tissues, cells, fluids Hand (Deceased-donor only) Cornea (Deceased-donor only) Skin including Face replant (autograft) and Face transplant (extremely rare) Islets of Langerhans (Pancreas Islet Cells) (Deceased-donor and Living-Donor) Bone marrow /Adult stem cell (Living-Donor and Autograft)

5 Blood transfusion /Blood Parts Transfusion (Living-Donor and Autograft) Blood vessels (Autograft and Deceased-Donor) Heart valve (Deceased-Donor, Living-Donor and Xenograft[Porcine/bovine]) Bone (Deceased-Donor and Living-Donor) Heart Transplant- is a surgical transplant procedure performed on patients with endstage heart failure or severe coronary artery disease. As of 2008 the most common procedure was to take a working heart from a recently deceased organ donor and implant it into the patient. The patient's own heart is either removed or, less commonly, left in place to support the donor heart both were controversial solutions to an enduring human ailment. Post-operation survival periods averaged 15 years. Heart transplantation is not considered to be a cure for heart disease, but a life-saving treatment intended to improve the quality of life for recipients. Lung Transplant- is a surgical procedure in which a patient's diseased lungs are partially or totally replaced by lungs which come from a donor. While lung transplants carry certain associated risks, they can also extend life expectancy and enhance the quality of life for endstage pulmonary patients. Kidney Transplant- a kidney transplant is an operation that places a healthy kidney in your body. The transplanted kidney takes over the work of the two kidneys that failed, so you no longer need dialysis. During a transplant, the surgeon places the new kidney in your lower abdomen and connects the artery and vein of the new kidney to your artery and vein. Often, the new kidney will start making urine as soon as your blood starts flowing through it. But sometimes it takes a few weeks to start working. Many transplanted kidneys come from donors who have died. Some come from a living family member. The wait for a new kidney can be long. If you have a transplant, you must take drugs for the rest of your life, to keep your body from rejecting the new kidney. Liver Transplant- is the replacement of a diseased liver with a healthy liver from another person. The most commonly used technique is orthotopic transplantation, in which the native liver is removed and replaced by the donor organ in the same anatomic location as the original liver. Liver transplantation is a viable treatment option for end-stage liver disease and acute liver failure. Typically three surgeons and two anesthesiologists are involved, with up to four supporting nurses. The surgical procedure is very demanding and ranges from 4 to 18 hours depending on outcome. Numerous anastomoses and sutures, and many disconnections and reconnections of abdominal and hepatic tissue, must be made for the transplant to succeed, requiring an eligible recipient and a well-calibrated live or cadaveric donor match. Intestine Transplant- is a last-resort treatment option for patients with intestinal failure who develop life-threatening complications from total parenteral nutrition (TPN). Long-term TPN can result in complications including bone disorders, catheter-related infections and liver failure. Over an extended period of time, TPN also can damage veins used to administer the nutrition via the catheter. Intestinal transplant is a complex procedure requiring a highly skilled multidisciplinary transplant team. An isolated Intestinal transplant surgery takes approximately three to four hours to complete whereas a multivisceral (multi-organ) transplant operation can take up to twelve hours.

6 Pancreas Transplant- is an organ transplant that involves implanting a healthy pancreas (one that can produce insulin) into a person who usually has diabetes. Because the pancreas is a vital organ, performing functions necessary in the digestion process, the recipient's native pancreas is left in place, and the donated pancreas is attached in a different location. In the event of rejection of the new pancreas which would quickly cause life-threatening diabetes, the recipient could not survive without the native pancreas still in place. The healthy pancreas comes from a donor who has just died or it may be a partial pancreas from a living donor. At present, pancreas transplants are usually performed in persons with insulin-dependent diabetes, who can develop severe complications. Patients with pancreatic cancer are not eligible for valuable pancreatic transplantations, since the condition has a very high mortality rate and the disease, being highly malignant, could and probably would soon return. Corneal Transplant- also known as corneal grafting, is a surgical procedure where a damaged or diseased cornea is replaced by donated corneal tissue in its entirety or in part. The graft is taken from a recently deceased individual with no known diseases or other factors that may affect the viability of the donated tissue or the health of the recipient. Skin Transplant- a skin graft is a surgical procedure that involves removing skin from one part of your body (the donor site) and moving it, or transplanting it, to a different part. This surgery may be done if part of your body has lost its protective covering of skin due to injury or illness. Blood Transfusion- the process of receiving blood products into one's circulation intravenously. Transfusions are used in a variety of medical conditions to replace lost components of the blood. Early transfusions used whole blood, but modern medical practice commonly uses only components of the blood, such as red blood cells, white blood cells, plasma, clotting factors, and platelets. Bone Transplant- transplants bone tissue. Surgeons use bone grafts to repair and rebuild diseased bones in your hips, knees, spine, and sometimes other bones and joints. Grafts can also repair bone loss caused by some types of fractures or cancers. Once your body accepts the bone graft, it provides a framework for growth of new, living bone. If the transplanted bone comes from another person, it is called an allograft. Most allograft bone comes from donors who have died. Tissue banks screen these donors and disinfect and test the donated bone to make sure it is safe to use. If the transplanted bone comes from another part of your own body, it is called an autograft. Autograft bone often comes from your ribs, hips or a leg.

TYPES OF DONOR LIVING- In living donors , the donor remains alive and donates a renewable tissue, cell, or fluid (e.g. blood, skin); or donates an organ or part of an organ in which the remaining organ can regenerate or take on the workload of the rest of the organ (primarily single kidney donation, partial donation of liver, small bowel). Regenerative medicine may one day allow for laboratory-grown organs, using patient's own cells (stem cells, or healthy cells extracted from the failing organs.)

7 DECEASED - (formerly cadaveric) are donors who have been declared brain-dead and whose organs are kept viable by ventilators or other mechanical mechanisms until they can be excised for transplantation. Apart from brain-stem dead donors, who have formed the majority of deceased donors for the last twenty years, there is increasing use of Donation after Cardiac Death - DCD- Donors (formerly non-heart beating donors) to increase the potential pool of donors as demand for transplants continues to grow. These organs have inferior outcomes to organs from a brain-dead donor; however given the scarcity of suitable organs and the number of people who die waiting, any potentially suitable organ must be considered.

LAW OF TRANSPLANTING REPUBLIC ACT NO. 7170- AN ACT AUTHORIZING THE LEGACY OR DONATION OF ALL OR PART OF A HUMAN BODY AFTER DEATH FOR SPECIFIED PURPOSES. The death of the person shall be determined in accordance with the acceptable standards of medical practice and shall be diagnosed separately by the attending physician and another consulting physician, both of whom must be appropriately qualified and suitably experienced in the care of such parties. The death shall be recorded in the patient's medical record. Living related donors- donors who have much more control over the timing of the transplant with a living donation. This may allow the transplant to occur before the recipient requires dialysis. Paired exchange- with paired-organ donation, your donor exchanges his or her kidney with the living donor from another incompatible donor/recipient pair to create two compatible pairs. While its true that your donor will not directly donate his or her kidney to you, exchanging with another incompatible pair will allow for two compatible transplants. Good Samaritan- is donors who do not know the recipient, but make their donation purely out of selfless motives. This type of donation is also referred to as anonymous or non-directed donation. Recipients are those at the top of the local wait list. Forced Donation- is the process when a person forced to donate a part/s or organ/s of another person. GENETIC ENGINEERING Genetic engineering, also called genetic modification, is the direct manipulation of an organism's genome using biotechnology. New DNA may be inserted in the host genome by first isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing the DNA, and then inserting this construct into the host organism. Genes may be removed, or "knocked out", using a nuclease. Glofish, the first GMO designed as a pet, was first sold in the United States December in 2003.

8 Process The first step is to choose and isolate the gene that will be inserted into the genetically modified organism. The gene can be isolated using restriction enzymes to cut DNA into fragments and gel electrophoresis to separate them out according to length. Polymerase Chain Reaction (PCR) can also be used to amplify up a gene segment, which can then be isolated through gel electrophoresis. If the chosen gene or the donor organism's genome has been well studied it may be present in a genetic library. If the DNA sequence is known, but no copies of the gene are available, it can be artificially synthesized. The gene to be inserted into the genetically modified organism must be combined with other genetic elements in order for it to work properly. The constructs are made using recombinant DNA techniques, such as restriction digests, ligations and molecular cloning. INVITRO FERTILIZATION Recombinant DNA technology or Genetic Engineering is a method that allows the combination of genes in a test tube to form a hybrid DNA. It allows the transfer of a specific gene (from the same or different organism) to produce a new trait in an organism. The true fathers of genetic engineering were American biochemists Stanley Cohen and Herbert Boyer, who were the first scientists to use restriction enzymes to produce a genetically modified organism. In 1973 they used type II enzymes to cut DNA into fragments, recombine the fragments in vitro, and then insert the foreign genes into a common laboratory strain of bacteria. The foreign genes replicated along with the bacteria's genome; furthermore, the modified bacteria produced the proteins specified by the foreign DNA. The new age of biotechnology had begun.

In Vitro Fertilization
Definition In vitro fertilization (IVF) is a procedure in which eggs (ova) from a woman's ovary are removed. They are fertilized with sperm in a laboratory procedure, and then the fertilized egg (embryo) is returned to the woman's uterus. In Vitro is used to refer to any biological procedure that is performed outside the organism, it would normally be occurring in to distinguish it from an In Vivo procedure, where the tissue remains inside the living organism within is normally found. A colloquial term for babies conceived as the result of In Vitro Fertilization (IVF), test tube babies, refers to the tube-shaped containers of glass or plastic resin, called test tubes that are commonly used in chemistry labs. However, In Vitro Fertilization is usually performed in the shallower container called Petri dishes. In Vitro Fertilization (IVF) is a process by which an egg is fertilized by sperm outside the body. In Vitro Fertilization is a major treatment for infertility when other methods of assisted reproductive technology have failed. The term in vitro, is from the Latin word meaning in glass, is used, because early biological experiments involving cultivation of tissues outside the living organisms from which they came were carried out in glass containers such as beakers, test tubes, or Petri dishes.

9 PROCESS The process of In Vitro Fertilization involves monitoring and stimulating a womans ovaries. A woman may be given fertility drugs before this procedure so that several eggs mature in the ovaries at the same time. Eggs (ova) are removed from a woman's ovaries using a long, thin needle. The physician gains access to the ovaries using one of two possible procedures. The first step in IVF involves injecting hormones so you produce multiple eggs each month instead of only one. You will then be tested to determine whether you're ready for egg retrieval. Prior to the retrieval procedure, you will be given injections of a medication that ripens the developing eggs and starts the process of ovulation. Timing is important; the eggs must be retrieved just before they emerge from the follicles in the ovaries. If the eggs are taken out too early or too late, they won't develop normally. Your doctor may do blood tests or an ultrasound to be sure the eggs are at the right stage of development before retrieving them. The IVF facility will provide you with special instructions to follow the night before and the day of the procedure. IVF is one of several assisted reproductive techniques (ART) used to help infertile couples to conceive a child. If after one year of having sexual intercourse without the use of birth control a woman is unable to get pregnant, infertility is suspected. Some of the reasons for infertility are damaged or blocked fallopian tubes, hormonal imbalance, or endometriosis in the woman. In the man, low sperm count or poor quality sperm can cause infertility. IVF is one of several possible methods to increase the chance for an infertile couple to become pregnant. Its use depends on the reason for infertility. IVF may be an option if there is a blockage in the fallopian tube or endometriosis in the woman or low sperm count or poor quality sperm in the man. There are other possible treatments for these conditions, such as surgery for blocked tubes or endometriosis, which may be tried before IVF. IVF will not work for a woman who is not capable of ovulating or a man who is not able to produce at least a few healthy sperm. Precautions The screening procedures and treatments for infertility can become a long, expensive, and sometimes, disappointing process. Each IVF attempt takes at least an entire menstrual cycle and can cost $5,000-$10,000, which may or may not be covered by health insurance. The anxiety of dealing with infertility can challenge both individuals and their relationship. The added stress and expense of multiple clinic visits, testing, treatments, and surgical procedures can become overwhelming. Couples may want to receive counseling and support. Description In vitro fertilization is a procedure where the joining of egg and sperm takes place outside of the woman's body. A woman may be given fertility drugs before this procedure so that several eggs mature in the ovaries at the same time. Eggs (ova) are removed from a woman's ovaries using a long, thin needle. The physician gains access to the ovaries using one of two possible procedures. One procedure involves inserting the needle through the vagina (transvaginally). The physician guides the needle to the location of the ovaries with the help of an ultrasound machine. In the other procedure, called laparoscopy, a small thin tube with a viewing lens is inserted through an incision in the navel. This allows the physician to see inside the patient, and locate the ovaries, on a video

10 monitor.Once the eggs are removed, they are mixed with sperm in a laboratory dish or test tube. (This is where the term test tube baby comes from.) The eggs are monitored for several days. Once there is evidence that fertilization has occurred and the cells begin to divide, they are then returned to the woman's uterus. In the procedure to remove eggs, enough may be gathered to be frozen and saved (either fertilized or unfertilized) for additional IVF attempts. A 2004 study from the Mayo Clinic found that frozen sperm was as effective as fresh sperm for IVF. Other types of assisted reproductive technologies might be used to achieve pregnancy. A procedure called intracytoplasmic sperm injection (ICSI) uses a manipulation technique that must be performed using a microscope to inject a single sperm into each egg. The fertilized eggs can then be returned to the uterus, as in IVF. In gamete intrafallopian tube transfer (GIFT) the eggs and sperm are mixed in a narrow tube and then deposited in the fallopian tube, where fertilization normally takes place. Another variation on IVF is zygote intrafallopian tube transfer (ZIFT). As in IVF, the fertilization of the eggs occurs in a laboratory dish. And, similar to GIFT, the embryos are placed in the fallopian tube (rather than the uterus as with IVF). TWO PROCEDURES OF IVF One procedure involves inserting the needle through the vagina called transvaginally. The physician guides the needle to the location of the ovaries with the help of an ultrasound machine. In the other procedure, called laparoscopy, a small thin tube with a viewing lens is inserted through an incision in the navel. This allows the physician to see inside the patient, and locate the ovaries, on a video monitor. EGG AND SPERM PREPARATION In the laboratory, the identified eggs are stripped of sorrounding cells and prepared for fertilization. An oocyte selection may be performed prior to fertilization to select eggs with optimal chances of successful pregnancy. In th meantime, semen is prepared for fertilization by removing inactive cells and seminal fluid in a process called sperm washing. If semen is being provided by a sperm donor, it will usually have been prepared for treatment before being frozen and quarantined, and it will be thawed ready for use. Preparation Once a woman is determined to be a good candidate for in vitro fertilization, she will generally be given "fertility drugs" to stimulate ovulation and the development of multiple eggs. These drugs may include gonadotropin releasing hormone agonists (GnRHa), Pergonal, Clomid, or human chorionic gonadotropin (hcg). The maturation of the eggs is then monitored with ultrasound tests and frequent blood tests. If enough eggs mature, the physician will perform the procedure to remove them. The woman may be given a sedative prior to the procedure. A local anesthetic agent may also be used to reduce discomfort during the procedure. The first successful birth of a test tube baby is Louise Brown. Louise Brown was born as a result of natural cycle In Vitro Fertilization where no stimulation was made. Robert G. Edwards, the physiologist who develop the treatment, was awarded the Nobel Prize in Physiology or Medicine in 2010.

11 On July 25, 1978, Louise Joy Brown, the world's first successful "test-tube" baby was born in Great Britain. Though the technology that made her conception possible was heralded as a triumph in medicine and science, it also caused many to consider the possibilities of future ill-use. CELEBRITIES W IVF KIDS 1.Celine Dion After six in vitro fertilization attempts, Celine Dion and her husband Rene Anglil gave birth to their son Ren-Charles in 2000. A decade later, the couple once again used IVF to conceive their twin boys Eddy and Nelson, born in 2010. IVF is difficult emotionally, Dion told Hello! Canada magazine. 2.Mariah Carey The chart-topping singer, who had previously suffered a miscarriage, underwent IVF treatments to get pregnant. In 2011, Carey and her husband Nick Cannon welcomed their twins Moroccan and Monroe, which she famously nicknamed "Dem Babies." 3.Nicole Kidman Nicole Kidman went through various IVF and fertility treatments before the 2008 birth of her and Keith Urban's daughter Sunday Rose. The couple's second child Faith Margaret was born in 2010 via a gestational surrogate 4.Courteney Cox After struggling to conceive, Courteney Cox and David Arquette's daughter Coco was born in 2004, thanks to IVF. 5.Brooke Shields Brooke Shields reportedly underwent seven IVF treatments before giving birth to her and Chris Henchy's daughter Rowan Francis in May 2003. The couple was able to naturally conceive their second daughter Grier Hammon three years later.

Aftercare After the IVF procedure is performed the woman can resume normal activities. A pregnancy test can be done approximately 12-14 days later to determine if the procedure was successful. Risks The risks associated with in vitro fertilization include the possibility of multiple pregnancy (since several embryos may be implanted) and ectopic pregnancy (an embryo that implants in the fallopian tube or in the abdominal cavity outside the uterus). There is a slight risk of ovarian rupture, bleeding, infections, and complications of anesthesia. If the procedure is successful and pregnancy is achieved, the pregnancy would carry the same risks as any pregnancy achieved without assisted technology. Normal results Success rates vary widely between clinics and between physicians performing the procedure and implantation does not guarantee pregnancy. Therefore, the procedure may have to be repeated

12 more than once to achieve pregnancy. However, success rates have improved in recent years, up from 20% in 1995 to 27% in 2001. Abnormal results An ectopic or multiple pregnancy may abort spontaneously or may require termination if the health of the mother is at risk. The number of multiple pregnancies has decreased in recent years as technical advances and professional guidelines have led to implanting of fewer embryos per attempt. Resources CLONING Cloning is the creation of an organism that is an exact genetic copy of another. This means that every single bit of DNA is the same between the two! TWO WAYS OF CLONING Artificial Embryo Twinning Artificial embryo twinning is the relatively low-tech version of cloning. As the name suggests, this technology mimics the natural process of creating identical twins. Somatic Cell Nuclear Transfer Somatic cell nuclear transfer, (SCNT) uses a different approach than artificial embryo twinning, but it produces the same result: an exact clone, or genetic copy, of an individual. This was the method used to create Dolly the Sheep. SCNT means Somatic cell: A somatic cell is any cell in the body other than the two types of reproductive cells, sperm and egg. Sperm and egg are also called germ cells. In mammals, every somatic cell has two complete sets of chromosomes, where as the germ cells only have one complete set. Nuclear: The nucleus is like the cell's brain. It's an enclosed compartment that contains all the information that cells need to form an organism. This information comes in the form of DNA. It's the differences in our DNA that make each of us unique. Transfer: Moving an object from one place to another. To make Dolly, researchers isolated a somatic cell from an adult female sheep. Next, they transferred the nucleus from that cell to an egg cell from which the nucleus had been removed. After a couple of chemical tweaks, the egg cell, with its new nucleus, was behaving just like a freshly fertilized zygote. It developed into an embryo, which was implanted into a surrogate mother and carried to term. The lamb, Dolly, was an exact genetic replica of the adult female sheep that donated the somatic cell nucleus to the egg. She was the first-ever mammal to be cloned from an adult somatic cell.

13 CLONING PROCESS OF DOLLY

TYPES OF CLONING REPRODUCTIVE Cloning which are used for producing the exact copy of an existing organism are called as reproductive cloning. Reproductive cloning is performed using a technique known as somatic cell nuclear transfer. During this procedure nucleus is extracted from a somatic cell. Somatic cell is any cell from the body other than the germ cell. Then the nucleus of the egg is also removed, the extracted nucleus is then transferred into enucleated egg. Egg starts to divide after receiving the external stimuli. Egg will divide and grow and finally will develop into an embryo. This embryo is then implanted into a gestational surrogate mother. Reproductive cloning has some issues like; clones which are produced have shorter lifespan when compared with the parent species. Reproductive cloning also imposes the risk of losing genetic diversity of the nature. THERAPEUTIC Biotechnology cloning used for the purpose of medical treatment are known as therapeutic cloning. Therapeutic cloning is mainly used to grow organs, which can then be used to replace the non functional organs of a patient. It can also be used to generate the skin to treat burn victims, can be used to create nerve cells to treat patients suffering from nerve degenerative diseases. Therapeutic cloning like reproductive cloning is also done by using somatic cell nuclear transfer techniques. The main difference is that the dividing egg is used to extract stem cells. Extracted stem cells are then used to produce required organs, by growing them in the suitable media.

14 Therapeutic cloning plays a major role in medical treatment. Doctors with the help of therapeutic cloning technique can grow replacement organs for damaged body parts of the patients. These organs are genetically and immunologically same as the patient's own organ. Therefore it reduces the usage of immunosuppressant drugs and also the risk of organ rejection by the body. DNA CLONING Biotechnology cloning techniques used to create many copies of DNA fragments are called as DNA cloning. This technique can be used to clone random fragment of DNA, specific sequence of DNA or can be used to clone entire gene sequence. DNA cloning process comprises many steps; first the interested DNA fragment is extracted from the genome, by the help of restriction enzymes or can be synthesized chemically. Polymerase chain reaction or cloning vectors are used to produce multiple copies of DNA fragments. Then agarose gel electrophoresis and DNA sonication methods are also used for cloning DNA fragments. DNA cloning is used in genetic engineering to create microorganisms, plants or animals with desirable features. DNA cloning is also used in DNA fingerprinting and also in gene sequencing. CLONED ORGANISMS The world's first cloned mule, was born on May 4. He is an identical genetic copy of his brother, a champion racing mule called Taz, and the first clone to be born in the equine family. The world's first cloned kitten, named Cc. It was created by scientists in Texas using a cell taken from an adult tortoiseshell female. The photo, taken on December 22 2001 when the kitten was seven weeks old, was made public in February 2002 Rainbow, the adult tortoiseshell female from which Cc was cloned. The nuclear-donor cat was used in the transfer technique pioneered by the Edinburgh scientists who made Dolly the sheep. The move opens the prospect of people being able to clone their pets. Five cloned female piglets, named Noel, Angel, Star, Joy and Mary - an important step towards "knock-out pigs". They were born on Christmas Day 2001 in what the Scottish-based firm PPL Therapeutics says is a major step towards successfully producing animal organs and cells for use in human transplants. The pigs lack a gene to which the human immune system reacts aggressively. When an all-male litter is born and bred with the females, a true knock-out pig will be created. Dolly the sheep, the world's first cloned adult animal. The scientists who cloned Dolly are to stop experiments involving genetically modifying pigs for human organ transplants because of concerns that deadly new diseases could be passed on to people. A pair of new-born cloned calves in a cowshed in Ishikawa Japan, on July 5 1998. They were born exactly two years after Dolly, the British sheep that made history by becoming the first clone of an adult animal. They are the second adult-animal clones, and were produced by a similar

15 technique. A spokesman for the Ishikawa prefectural livestock research centre said the new technique would be used to breed better cattle strains with higher-quality beef or greater milk capacity. A pair of new-born cloned calves in a cowshed in Ishikawa Japan, on July 5 1998. They were born exactly two years after Dolly, the British sheep that made history by becoming the first clone of an adult animal. They are the second adult-animal clones, and were produced by a similar technique. A spokesman for the Ishikawa prefectural livestock research centre said the new technique would be used to breed better cattle strains with higher-quality beef or greater milk capacity. ANDi (inserted DNA spelled backwards), the first genetically modified rhesus monkey, at the Oregon regional primate research centre in Oregon, USA. The birth of ANDi, the first rhesus monkey cloned by embryo splitting, is another incremental step toward designing and perfecting new treatments for human genetic disorders. GENOME MAPPING A genome is the entirety of an organism's hereditary information. It is encoded either in DNA or, for many types of viruses, in RNA.

The Human Genome Project (HGP) was one of the great feats of exploration in history an inward voyage of discovery rather than an outward exploration of the planet or the cosmos; an international research effort to sequence and map all of the genes - together known as the genome - of members of our species, Homo sapiens. GENOME MAPPING Genetic mapping - can offer firm evidence that a disease transmitted from parent to child is linked to one or more genes. It also provides clues about which chromosome contains the gene and precisely where it lies on that chromosome. Gene mapping, also called genome mapping, is the creation of a genetic map assigning DNA fragments to chromosomes. Locating of a specific gene to particular region of a chromosome and determining the location of and relative distances between genes on the chromosome. To produce a genetic map, researchers collect blood or tissue samples from family members where a certain disease or trait is prevalent. Using various laboratory techniques, the scientists isolate DNA from these samples and examine it for the unique patterns of bases seen only in family members who have the disease or trait. These characteristic molecular patterns are referred to as polymorphisms, or markers. When a genome is first investigated, this map is nonexistent. The map improves with the scientific progress and is perfect when the genomic DNA sequencing of the species has been completed. During this process, and for the investigation of differences in strain, the fragments are identified by small tags. These may be genetic markers or the unique sequence-dependent

16 pattern of DNA-cutting enzymes. The ordering is derived from genetic observations for these markers or in the second case from a computational integration of the fingerprinting data. The term "mapping" is used in two different but related contexts. GENETIC MARKERS Markers themselves usually consist of DNA that does not contain a gene, however they can tell a researcher the identity of the person a DNA sample came from.

GENOME MAP A genome map helps scientists navigate around the genome. Like road maps and other familiar maps, a genome map is a set of landmarks that tells people where they are, and helps them get where they want to go.

USES OF GENOME MAPPING Identify genes responsible for diseases. Heritable diseases Cancer

Identify genes responsible for traits. Plants or Animals Disease resistance Meat or Milk Production

TYPES OF MAP Nucleotide Sequence Maps complete or partially sequenced organisms

Cytogenetic Maps Breakpoints in disease Direct binding of probes to chromosome

Linkage Map It shows the arrangement of genes and genetic markers along the chromosomes as calculated by the frequency with which they are inherited together. Physical Map It is the representation of the chromosomes, providing the physical distance between landmarks on the chromosome, ideally measured in nucleotide bases. Physical maps can be

17 divided into three general types: chromosomal or cytogenetic maps, radiation hybrid (RH) maps, and sequence maps. TYPES OF PHYSICAL MAP Cytogenetic map is the visual appearance of a chromosome when stained and examined under a microscope. Particularly important are visually distinct regions, called light and dark bands, which give each of the chromosomes a unique appearance. This feature allows a person's chromosomes to be studied in a clinical test known as a karyotype, which allows scientists to look for chromosomal alterations. Radiation hybrid map uses X-ray breakage of chromosomes to determine the distances between DNA markers, as well as their order on the chromosome. In addition, the method allows the relative likelihoods of alternative marker orders to be determined. The RH procedure was used to map 14 DNA probes from a region of human chromosome 21 spanning 20 mega base pairs. Sequence Map has been established by analyzing the segregation of 288 sequencecharacterized genetic markers in an F(2) population composed of 93 individuals. GENETIC ENGINEERING Genetic engineering, also called genetic modification, is the direct manipulation of an organism's genome using biotechnology. It includes isolating, copying and multiplying genes, recombining genes or DNA from different species, and transferring genes from one species to another. The term Genetic Engineering" was first coined by Jack Williamson in his science fiction novel Dragon's Island; published in 1951. STEPS The first step is to choose and isolate the gene that will be inserted into the genetically modified organism. The promoter region initiates the transcription of the gene and can be used to control the location and level of gene expression, while the terminator region ends the transcription. The selectable marker, which in most cases confers antibiotic resistance to the organism it is expressed in, is needed to determine which cells are transformed with the new gene. The constructs are made using recombinant DNA techniques, such as restriction digests, ligations and molecular cloning. Genome editing is a type of genetic engineering in which DNA is inserted, replaced, or removed from a genome using artificially engineered nucleases, or "molecular scissors."

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