Reproduction and Fetal Technology

1. Reproduction and Fetal Technology Annabel Barajas Tamari Zeigler Thomas Ji Jared Salisbury Nathan Vasquez

2. Genetic Engineering and Gene Therapy • Both genetic engineering and gene therapy have the same basic premise: of having an introduction of a specific new genetic material into an organism’s genome

3. Some General Genetic Facts • A single chromosome contains about 1000 genes • Humans have approximately 30,000 different kinds of genes spread out over the 46 chromosomes • Humans get one complete set of genes from each parent

4. More General Facts on Genes and Disease Relationship’s • An abnormal gene is called a mutation • Abnormal genes can cause disease or , more commonly, can change your risk of developing a disease • Most common diseases are caused by a mixture of environmental, dietary and genetic factors • In a normal cell , it is common for many genes to be “ silent” (not expressed)

5. So what is genetic engineering? • Genetic engineering refers to the deliberate design of an entire organism. The process involves altering the genetic material of a single cell in a very small developing embryo, which then develops into an entire organism, with the altered genetic material in every cell. • The basic strategy is to place the new genetic material into the cell, and use the cell’s natural machinery to integrate this material into the genome

6. There are several ways to conduct the process of genetic engineering • One way is through horizontal gene transfer. Horizontal gene transfer refers to the transfer of genes or genetic material directly from one individual to another by processes similar to infection. • "Somatic" genetic engineering is genetic engineering that targets the genes in specific organs and tissues of the body of a single existing person without affecting genes in their eggs or sperm. Somatic gene transfer experiments are currently undergoing clinical trials, with mixed results to date. • "Germline" genetic engineering is genetic engineering that targets the genes in eggs, sperm, or very early embryos. The alterations affect every cell in the body of the resulting individual, and are passed on to all future generations.

7. The dangers, moral and ethical questions raised by genetic engineering • Genetic engineering is inherently hazardous, because it depends on designing artificial vectors to cross all species barriers, greatly increasing the potential for generating new viral and bacterial pathogens by horizontal gene transfer and recombination • Genetic engineering poses unprecedented ethical and social concerns, as well as serious challenges to the environment, human health, animal welfare, and the future of agriculture • Also concerns about the consequences of altering a few select genes, since genes interact in very complex ways, and we may not be able to predict all the consequences of any specific manipulation.

8. So what is gene therapy? And how does it work? • Gene therapy is an experimental technique that uses genes to treat or prevent disease • In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patient's cells instead of using drugs or surgery • Gene therapy is designed to introduce genetic material into cells to compensate for abnormal genes or to make a beneficial protein • If a mutated gene causes a necessary protein to be faulty or missing, gene therapy may be able to introduce a normal copy of the gene to restore the function of the protein.

9. A Diagram showing the process

10. A genome- is the totality of the genetic material of a cell or organism • Vector - A vector is a small piece of DNA used to carry a gene of interest. Besides the gene being studied, a vector may contain elements which are used to help the gene integrate into a genome • Recombination - The process in which strands of DNA are cut open and rejoined. This is done by a complex of proteins called DNA recombinase. Recombination can be used to integrate the DNA from a vector into an organism's genome

11. Is gene therapy safe? • Gene therapy is under study to determine whether it could be used to treat disease • Current research is evaluating the safety of gene therapy; future studies will test whether it is an effective treatment option. Several studies have already shown that this approach can have very serious health risks, such as toxicity, inflammation, and cancer • Because the techniques are relatively new, some of the risks may be unpredictable; however, medical researchers, institutions, and regulatory agencies are working to ensure that gene therapy research is as safe as possible.

12. What are the ethical issues surrounding gene therapy? • How can "good" and "bad" uses of gene therapy be distinguished? • Who decides which traits are normal and which constitute a disability or disorder? • Will the high costs of gene therapy make it available only to the wealthy? • Could the widespread use of gene therapy make society less accepting of people who are different? • Should people be allowed to use gene therapy to enhance basic human traits such as height, intelligence, or athletic ability? • The idea of germline gene therapy is controversial. While it could spare future generations in a family from having a particular genetic disorder, it might affect the development of a fetus in unexpected ways or have long-term side effects that are not yet known. • Because people who would be affected by germline gene therapy are not yet born, they can't choose whether to have the treatment. Because of these ethical concerns, the U.S. Government does not allow federal funds to be used for research on germline gene therapy in people

13. Artificial Insemination-Intrauterine Insemination (IUI) • “Granddaddy” of fertility treatments. It dates back to the early 1900’s. • “First Line” treatment used before any other Assisted Reproductive Technologies (ART’s) are used. • It is used when an unexplained fertility problem is present. • Very simple procedure: only healthy sperm, and delivery device, and a female to receive the sperm are needed. • Much less invasive than other ART procedures

14. Process • Female usually undergoes a cycle (at least one week) of Clomid or FSH to stimulate development of eggs. • Once ovulation occurs a sample of sperm is collected either from father or from a sperm bank. • The sperm is then “washed.” Which concentrates the hardiest sperm and removes any potential antigens that may cause allergic reactions in the female. • The “washed” sperm in delivered directly to the uterus using a catheter. • Two weeks later a simple over the counter pregnancy test will determine if the female is pregnant or not.

15. Advantages • -If the female has a sperm allergy • -If the male has a low sperm count or poor sperm mobility • -It allows fertilization to occur naturally inside the body unlike other ART’s • -If antisperm anti bodies are present in vagina they are bypassed using catheter • -No father is needed, can acquire sperm from sperm banks. • -Cost: average ranges between $300-$700 per cycle

16. Disadvantages • -Most couples only have a 5%-25% success rate using IUI • -May have to repeat process several times to achieve pregnancy • -Weak children may be a result due to the fact that weaker sperm have a chance to reach the egg.

17. Human Cloning • Cloning is the reproduction of an individual single cell taken from a donor or parent.

18. Somatic Cell Nuclear Transfer (Therapeutic Cloning) • Egg from female donor is taken and the nucleus is removed creating an enucleated egg. • A somatic cell (non-reproductive cell) is then fused together with the enucleated egg producing an embryo. • The embryo is implanted into the surrogate mother using in vitro fertilization. • If it is a success, nine months later an exact clone of the somatic cell donor is created.

19. After hundreds of attempts Dolly the sheep was created. After a short life of 6 years she was put to sleep after suffering from a progressive lung disease.

20. Questions raised: • Scientists are concerned that the clone will have cells that are the same age as the donor cells. • Will parents try to live the life they weren’t able to through their exact clone? • Will knowing that they are not only a child but also a sister/brother of parents emotionally damage the clone?

21. Assisted Reproductive Technology (ART) • Most ART procedures are recommended in cases of ovulation problems, blocked fallopian tubes, low sperm count in partner, and if other treatment method are unsuccessful. • Doctor prescribes fertility drugs prior to ART procedures to stimulate ovaries to develop several mature eggs for fertilization. The doctor will then monitor blood hormone levels to detect when the eggs are mature. • Since the sperm is put in the direct vicinity of the eggs in a dish it is possible to achieve fertilization with a lower than average sperm count. • Since more than one embryo may be implanted there is a higher possibility of multiple birth pregnancies than with natural child birth.

22. In Vitro Fertilization (IVF) • A procedure in which a sperm and oocyte are combined in a laboratory dish and implanted in the uterus. • IVF is the most common ART procedure used, accounting for more than 70% of ART procedures. • In the past eggs were retrieved using a laparoscopy procedure; however, due to advancements in technology eggs are now retrieved vaginally with the use of an ultrasound to determine the location of the eggs. • After about 2-5 days the fertilized eggs form an embryo and are then implanted in the uterus, using a catheter, to develop naturally. • Treatment takes about six weeks to complete.

23. Gamete Intrafallopian Transfer (GIFT) • A procedure similar to IVF but in GIFT the eggs and sperm are transferred into the fallopian tubes immediately after the two are mixed. There is no time given for the eggs to be fertilized before the transfer. • GIFT accounts for about 2 percent of ART procedures. • The egg and sperm mixture are implanted in the fallopian tubes using a laparoscopy procedure. • One cycle of GIFT takes about six weeks. • GIFT is the closest ART procedure to natural childbirth.

24. Zygote Intrafallopian Transfer (ZIFT) • Similar to GIFT except the egg is fertilized before it is placed in the fallopian tubes. • ZIFT also referred to as tubal embryo transfer (TET). • It is different from IVF since the embryo is placed in the fallopian tubes instead of the uterus. • Only accounts for one percent of ART procedures. • Zygotes are transplanted about one day after fertilization. • If the treatment is successful the zygote will travel to the uterus and implant itself, where it will develop.

25. Frozen Embryo Transfer (FET) • Frozen embryos can be frozen after 24-72 hours in a culture. • Frozen embryos can be stored for several years. • Embryos will be thawed and replaced in the uterus once ovulation is identified by a doctor or when the endometrium is thick enough. • Not all embryos survive the freezing, storage, and thawing processes. • Thawed embryos are implanted in the uterus using a catheter.

26. Intracytoplasmic Sperm Injection (ICSI) • This procedure was introduced in 1982, and continues to be a popular treatment in treating male infertility. • ICSI is prescribed in cases of low sperm count, missing vas deferens, or irreversible vasectomy. • The male provides the doctor with a sperm sample by ejaculation. However, if there is not enough sperm in ejaculate the doctor may remove sperm from the testicles using a needle or perform a biopsy of the testicular tissue. • There is concern for the long term health and development of children conceived by ICSI since it enables weaker sperm to fertilize eggs.

27. Surrogate Motherhood • Surrogate mothers are women who carry a child for a mother who is unable to conceive with the intention of giving the child up once it is born. • Also known as ersatz mothers • Surrogate pregnancies are conceived through artificial insemination. • In gestational surrogacy the surrogate mother is implanted with the real parent’s embryo using an IVF procedure. • Some gay male couples use surrogate mothers to have children that are biologically related to at least one of the partners. • Surrogate mothers usually receive compensation for carrying the child. • There are many ethical and legal concerned with this process since some surrogate mothers grow attached to the fetus and sue for custody of the child they carried. The most publicized case is that of Baby M.

28. Storage of gametes and embryos – Sperm cryopreservation & embryo cryopreservation • Gamete is a sex cell containing the genetic material necessary for reproduction • Embryo is the early form of life in the uterus between the stages of blastocyst and fetus • Cryopreservation is a process to store gametes or embryos by freezing them at low temperatures

29. Sperm cryopreservation & Embryo cryopreservation • Collect semen from self-masturbation • More portions of semen can be cryopreserved. This so-called “test thaws” will be warmed up to verify success • Retrieve egg under ultrasound guidance • Embryo can be frozen at any stage • Both sperm and embryo are stored in liquid nitrogen, around -196C

30. Benefits from Sperm & Embryo cryopreservation • People facing surgery, radiation therapy or chemotherapy for cancer or other medical operations that can make them sterile • If a male partner is expected to be away, cryopreserved sperm can be used in time for optimal insemination • Cryopreservation can help people with internal problem such as cervical stenosis • Frozen sperm or embryos can be donated to infertile couples • Lesbian couples who wants a child from cryopreservation method if they don’t desire sexual relationship with a male partner

31. Problems of Sperm & Embryo Cryopreservation • Not all sperms and embryos can survive the freezing and thawing process • Pregnancy rate using frozen sperm or embryo may have lower chance compared to using fresh sperm or embryo • No apparent birth defects from using frozen-thawed sperm but it is not sure whether problems will not appear in the future • Chance of possible infection with a bacteria or virus

32. Choices and Moral Issues • Couples have choices to: • continue to store them • arrange for the transfer treatment cycle before expiration • discard them by interfering during the thawing process • donate them to approved research facility or to infertile couple. • Most countries have different policies on how long sperm & embryo can be stored, United Kingdom is 10 years • Moral issues surrounding the cryopreservation process: • the fate of cryopreserved sperm or embryo on the death of couple who preserved them • ownership of the sperm or embryos if the couple separates, safety of sperm & embryo cryopreservation • a possible detrimental effect on the outcome of cryopreserved sperm & embryo transfer and possible increase in fetal abnormalities after long period of cryopreservation.

33. Fertility Drug Treatments

34. WHAT ARE THEY FOR? • Women • Treat irregular ovulation • Help those who do not ovulate at all • Polycystic ovarian syndrome (PCOS) • Men with hormonal imbalances • Low sperm count • Poor sperm quality or motility • Primary hypogonadotrophic hypogonadism

35. TYPES • Clomiphene • Oral Pill • Gondotropins • Shot • Bromocriptine • Oral or Vaginal Pill

36. HOW LONG IS THE TREATMENT? • Clomiphene- You start 3 to 5 days into your cycle and continue for about 5 days • Gonadotrophins- 7 to 12 days each month • Bromocriptine- 2 to 3 times a day until you get pregnant

37. POSSIBLE SIDE EFFECTS • Bloating • Fluid retention • Weight gain • Nausea • Dizziness • Fatigue • Diarrhea • headaches • Abdominal tenderness • Stomach pain • Brest tenderness • Blurred vision • Irritability • Depression

38. CAN THEY CAUSE CANCER? • Beliefs • 1999 & 2000 studies

39. SUCCESS RATES It varies by treatment: • Clomiphene- 80 percent ovulate of this 40 percent get pregnant • Gonadotrophins- 20 to 60 percent conceive • Bromocriptine- 90 percent ovulate, 65 to 85 percent get pregnant • COST • Clomiphene- $50 on one cycle of Clomid • Gonadotrophins- $2,000 to $5,000 per cycle • Bromocriptine- $75 to $112 on one cycle * This is in the United States.

40. SEX SELECTION

41. QUICK FACTS • Males and Females • Carry 23 pairs of chromosomes • Difference between males and females • Lies in the 23rd chromosome • Girls have two X chromosomes • Boys have one X and one Y chromosome

42. IDENTIFYING THE SEX • Before implanting the embryo into the mother, a fertility doctor stains the chromosomes. • The x chromosome will be a different color than the Y chromosome. • Under the microscope, the doctor can easily tell whether the embryo is male or female.

43. THREE MAIN TECHNIQUESOF SEX SELECTION • Sperm Sorting Before Conception • Preimplantation Genetic Diagnosis (PGD) • Microsort

44. SPERM SORTING BEFORE CONCEPTION • Sperm sorting can be carried out either by putting live sperm in a dense liquid and spinning it in a centrifuge to separate sperm carrying the X chromosome and those carrying the Y chromosome.

45. PRE-IMPLANTATION GENETIC DIAGNOSIS • This procedure is carried out before the embryo is transferred into the woman. • A cell is removed from the embryo for genetic testing to detect X or Y chromosomes. • It is more controversial technique than sperm sorting because it involves embryos.

46. MICROSORT • This procedures involves using a fluorescent dye which binds to DNA in chromosomes in the semen sample, allowing the sperm to be separated using a laser.