EMBRYO TRANSFER TECHNOLOGY

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Embryo Transfer

Refers to a step in the process of assisted reproduction. The development of Embryo Transfer Technology (ETT) has emerged as a very Click to powerfuledit Master subtitle style insemination for tool after artificial genetic improvement programme. It is a complex process and involves multiple steps resulting in the insemination and fertilization of oocytes (eggs) in the laboratory . Used in connection with In vitro fertilization (IVF)), It may be used in humans or in animals, in 4/28/12

Refers to the technology by which fertilized ova are collected from the reproductive tract of genetically superior female (donor) and transferred to that of another female (recipient) which is genetically inferior. ETT now offers new opportunity to improve the dairy industry.

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Historical Perspective: Heape, student at Cambridge In 1890 Waiter

University performed the first embryo transfer experiment. He removed surgically two embryo (4-cell stage) from Angora rabbit and transferred it in a Belgian female hare. Afterwards the events occurring in different year are as follows: 1933:- J. S. Nicholson successfully transferred embryo in rat. 1951:- El willett usefully transferred embryo in cattle.
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1960:- Kvasnickni, a Russian scientist produced first successful embryo transplanted offspring in swine. 1964:- Mutter and co-workers transferred a bovine embryo non surgically and getting a viable offspring. 1979:- Steptoe and Edwards got success in getting a human baby girl through ETT. 1983:- Drost got calf born by ETT. 1991:- Mishra and coworker successfully produced calf from frozen thawed buffalo embryo.
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What is embryo transfer? Embryo transfer takes place after eggs have been collected and fertilised in the laboratory. Depending on your situation between one and three of the best quality embryos are selected and then transferred to the womans womb. An embryo must successfully attach itself to the wall of the womb for pregnancy to begin.
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Embryo Transfer - Is It Necessary?

It has enabled many infertile couples to experience the joy of parenthood. This is specifically used in the artificial insemination for both humans and animals alike. The process is called In Vitro Fertilization (IVF); this involves inserting multiple embryos in the uterus of the female with the purpose of triggering a successful pregnancy.

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Why is embryo transfer necessary?

There are many, many reasons why this process may be applied. In humans, this is a boon for infertile couples. This is specifically helpful when the sperm count is low and pregnancy cannot occur due to lack of contact between the egg and the sperm. the conditions of the woman's reproductive organs are such that they prevent fertilization though the eggs are okay.

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Ovulati on

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The Process Of Ovulation

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Video Illustrating the detailed process of Ovulation

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Designed to stimulate the ovaries to produce several eggs (oocytes) rather than the usual single egg as in a natural cycle. During the normal reproductive cycle, two hormones - follicle-stimulating hormone (FSH) and luteinizing hormone (LH) - are required to initiate and complete the process of egg maturation.

Superovula tion

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Medications are administered to women either orally or by injection to increase levels of both LH and FSH. The goal of ovulation induction (OI) is to grow and ovulate an egg in a women who normally does not ovulate, while the goal of superovulation (SO) is to produce more than one egg to improve fertility in a women

who

already ovulates.
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Commonly used fertility medications Clomiphene Citrate tablet brands are taken by mouth
usually at a dose of 50 mg daily for 5 days between days 5-9 of your menstrual cycle. In patients with shorter cycles the medication is sometimes started earlier on day 3 of cycle.

Serop Clo Injectable Gonadotropin pens are given to help a woman hene mid

produce and release healthy eggs when ovulation is a problem or to stimulate the ovaries to produce multiple eggs during In Vitro Fertilization (IVF)..

Follistim Gonal F
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HCG trigger an HCG medication given to "trigger"

ovulation when ultrasound shows that the largest (dominant) follicle is between 16-20 mm. The medication is sometimes utilized to maintain adequate progesterone production from the ovary after ovulation.

Progesterone preparations utilized to maintain

optimal progesterone levels after ovulation and through the first 12 weeks of pregnancy. The medications can be given in the form of vaginal tablets, vaginal suppositories or injections.

dometrin tablets

Crinone cream Progesterone injection 4/28/12

GnRH medications
GnRH agonists are used are given to suppress pituitary function during IVF. In the treatment of endometriosis or uterine fibroids. During IVF it important to prevent your own trigger to ovulation. They can be given beginning a week before your menses . They can also be given together with the gonadotropin injections beginning on day 2-3 of cycle (micro-dose or short protocol). GnRH antagonists such as Ganirelix or Cetrotide are given daily, beginning on day 6 of gonadotropin injections. Cetrotide
Lupr on

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What about the risks of gonadotropin therapy?

As with any medical treatment, there are some risks specific tofertility medications: Multiple pregnancy - (mostly twins) occur in

approximately 20% of ovulation induction cycles utilizing gonadotropin injections. Poor response - treatment is discontinued during IVF when there is no response or when less than 3 eggs develop. When this happens, a different stimulation protocol utilizing higher gonadotropin dose is usually advised. Cycle cancellation Gonadotropin injections are discontinued and ovulation is prevented when too many eggs develop or when the estrogen level is very high, as this may increase the risk of multiple pregnancies and 4/28/12

Ovarian Hyperstimulation Syndrome (OHSS) - is a potentially life threatening

complication resulting from overstimulation of the ovaries. The condition is associated with ovarian enlargement, torsion (twisting) of the ovaries, weight gain, accumulation of abdominal fluid, decrease of blood volume and low blood pressure,

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All these symptoms lead to hospitalization and aggressive treatment. OHSS may occur even with mild stimulation. Fortunately, the severe form of Ovarian Hyperstimulation Syndrome is uncommon, occurring in less than 1% of patients.

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Retrieving the Oocytes (egg retrieval)

Collection of eggs is performed under transvaginal ultrasound guidance. To accomplish this, a needle is inserted (under IV sedation) through the vaginal wall into the ovaries using ultrasound to locate each follicle. The follicular fluid is drawn up into a test tube to obtain the eggs. Although patients are given pain medications intravenously and are carefully monitored by an anesthesiology staff, Some women may experience some discomfort during the procedure. Generally, the oocyte (egg) retrieval takes 20-30 minutes. Patients are usually discharged home within hours after the retrieval. 4/28/12

Risks of oocyte (egg) retrieval may include, but are not limited to, the following:

Potential reactions from the drugs and procedures used in the administration of anesthesia. Risks associated with the passage of the needle through the vagina into the ovaries (including infection, bleeding etc..)

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Collecting and Preparing the Sperm:

A semen sample will be obtained from the partner by masturbation on the day of the oocyte (egg) retrieval. This is usually obtained while the retrieval is being performed. After the specimen is produced, the sperm will be prepared for inseminating the collected eggs in the laboratory. Men who feel that they may have difficulty producing a semen specimen have the opportunity to have their specimens frozen by the laboratory ahead of time for use in this situation. Testicular biopsy can also be performed as a method to extract sperm for IVF.

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Insemination of Eggs /Fertilization Process

Following egg retrieval, the follicular fluid is immediately transferred to the laboratory for identification of eggs, evaluation, and preparation for insemination. In the process of collecting the follicular fluid, it is possible that a large number of eggs may be retrieved. It is strongly recommended that all of these eggs be inseminated to maximize the number of embryos available for subsequent transfer. The prepared sperm is added to each egg and they will be allowed to incubate overnight under controlled laboratory conditions. The next day, each egg is evaluated for 4/28/12

However, it is possible that no eggs are fertilized. If this happens, the laboratory staff will re-inseminate the eggs or perform intracytoplasmic sperm injection (ICSI) in hopes of obtaining embryos for transfer If fertilization still does not occur, the eggs will be discarded and the remainder of the procedure will be cancelled. In the case of severe male factor, the couple may be asked to consider the option of using anonymous donor sperm (obtained through a licensed sperm bank for use as a "backup" or secondary sperm source) if it is not possible to obtain sufficient sperm from the partner at the 4/28/12 time of fertilization.

A fertilized egg is called an embryo. An embryo goes through stages of development until it is ready for transfer to the uterus.

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For the eggs that have fertilized

They will be allowed to develop for two or more additional days under controlled laboratory conditions before they are placed inside the woman's uterus. Depending upon the couple's wishes, some fertilized eggs/ embryos may be frozen and stored for future use. Embryos may be transferred to the patient on day 1(zygote stage) of development, on day two (two-cells to four-cells stage), on day three (six-cells to eight-cells stage), on day four (morula), or on days five to seven (different blastocyst stages)

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Growth of an embryo from the 2-cell stage to Blastocyst.

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In these pictures actual photographs of an embryo is shown under magnification:

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Three to five days following the Egg Retrieval the patient will have the Embryo Transfer (ET). During this time the fertilized eggs (embryos) have been allowed to grow and divide in the incubator. The patient would have also been started on Progesterone injections and/or suppositories the day of HCG injection to prepare the uterine lining for implantation.

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Transferring Embryos to the Uterus The embryo transfer typically takes place under sterile conditions, even though you will not be placed under anesthesia. Steps of Embryo transfer Include:1.

With the assistance of the embryologist, the embryos are loaded into a special catheter. A speculum is placed into the vagina to allow visualization of the cervix, which will then be cleaned. Under ultrasound guidance, the catheter is placed through the cervix and into the uterus.
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1.

1.

4.

When the catheter has been placed appropriately, the embryos are gently inserted into the uterus where they will hopefully implant. Typically, two to four embryos are be transferred in one treatment cycle.

5.

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The Process of embryo transfer Diagrammatic View

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Video Illustrating Embry Transfer Process

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Catheter(imag es)

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Speculum:-A speculum

is a medical tool for investigating body cavities, with a form dependent on the body cavity for which it is designed. Like an endoscope, a speculum allows entry into a body cavity; endoscopes, however, tend to have optics while a speculum is intended for direct vision.

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After the embryos are transferred to the womb, the woman will continue progesterone supplementation that begins on the evening of your egg retrieval procedure. Progesterone can be taken as a combination of oral troches and rectal/vaginal suppositories or by injections. Administration of these medications after egg collection has been shown to create a more

favorable uterine environment for the embryos, which therefore increases

pregnancy rates.

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Side effects of progesterone may include the following: Vaginal dryness; Bloating, Breast tenderness; Depression, Mood swings; Delay of menses

1. 2. 3. 4. 5. 6.

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Additional aspects of successful implantation include:

Bed rest following embryo transfer, The absence of hydrosalpinx (blocked, fluidfilled fallopian tubes) that could lead to washing out of the implanted embryos by intermittent leakage of the hydrosalpingeal fluid. If uterine contractions occur after transfer, the embryos could be expelled down through the cervix, or up into the fallopian tubes, instead of implanting in the uterus. Additionally, the presence of blood on the outside of the transfer catheter tip correlates with lower rates of successful implantation. 4/28/12

Risk of Embryo Transfer! Embryo transfer can cause:

Mild cramping, During the embryo transfer the embryo(s) may be displaced through the cervix (causing loss of embryos) or Into the fallopian tubes (causing possible tubal pregnancy). There is a small risk of bleeding or infection as a result of the transfer procedure.

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Interaction between the embryo and the endometrium Video Showing the implantation process

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Technique of Freezing Embryo

Cryopreservation. Crucial procedure for success of embryo transfer in livestock species. Here embryo is dehydrated with the help of a cryoprotectant like glycerol. After the equilibration is achieved embryos are cooled very slowly from 25C to - 38C. The embryo at -38C are plunged into liquid nitrogen (-196C) and stored for posterity and used whenever needed.
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Cryopreservation has the following advantages: It relieves for having simultaneous synchronization of oestrus in donor and recipient animals. Easy and safe transport of valuable germplasm throughout the globe. Conservation of superior genetic material. Protection of valuable strains of experimental animals against possible loss through disease, accident or genetic drift. Possibility of shortening the generation interval for progeny testing programme.
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according to our requirement.

Embryo Sexingdesired sex Embryo sexing enable us to have

Example - If we want animals for meat purposes with faster growth and lean carcasses then certainly we would like to have more males, but if we want animal for milk purposes than we will like to have more female for dairy purpose. Now these desired traits can be obtained by sexing embryo. There are two approaches for the control of sex of offspring: 1. regulation of sex of embryo 2. identification of sex of embryo. 4/28/12

 Regulation of Sex of Embryo are Based on: Sperm separation Nuclear transplantation Parthenogenesis and Removal of few cell from an embryo at an early stage of development and to make a preparation from these cells in which the chromosomes can be examined. Such removal of cells do not restrict its further developmental capacity.
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 Sperm separation

Sperm sorting is an advanced technique that sorts sperm "in vitro" byflow cytometry. This shines alaserat the sperm to distinguish X and Y chromosomes, and can automatically separate the sperm out into different samples. The technology is already in commercial use for animal farming.It is currently being trialed on humans in the US under the trademark MicroSort; it claims a 90% success rate but is still considered experimental by theFDA
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 Parthenogenesis

Parthenogenesisis a form ofasexual reproduction found infemales, where growth and development ofembryos occur withoutfertilization by a male. Inplants, parthenogenesis means development of an embryo from an unfertilized egg cell, and is a component process ofapomixis. The word "parthenogenesis" comes from theGreek,parthenos, meaning "virgin", and ,genesis, meaning "birth".The term is sometimes used inaccurately to describe reproduction modes inhermaphroditicspecies which can reproduce by themselves because they contain reproductive organs of both sexes in a single individual's body.
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Identification of Sex of Embryo:- This can be done by: Karyotyping-can be done by using quinacirine mustard. Identification of sex chromatin. Assay of sex linked enzymes, like glucose-6phosphate dehydrogenase. Identification of DNA probe or Y -chromosome and Serological determination of H-Y antigen which is a male specific antigen in mammals. These include attempts to develop monoclonal antibodies against H4/28/12 Y antigen and to detect sex by means of ELISA.

Micro-Manipulation of The technique is based on the developmental and Embryo

regulatory capacity of individual cells or groups of cells from embryo at early cleavage stage. By micromanipulation of embryo we can have Monozygotic twins in sheep, cattle, pigs and horses. This have been derived by separating the blastomeres from either two, four or eight-celled embryo into the equal groups. A large number of identical animals from a single embryo by the simple technique of separating blastomeres. (Single blastomere from 8-celled embryo or group of two blastomere from I6-celled embryo 4/28/12 seldom produce a viable foetus).

A set of identical quadruplet lambs derived from single 8-celled embryo.

Chimeric animals by this technique. Chimera is a composite animal living cells from more than one cell line. The main objective is to combine the best genetic traits of each donor in the chimeric animaL Sheep-goat chimera has been successfully produced.

Many exciting possibilities for manipulation of animals of selected genotype.

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Advantages of Embryo Transfer Technology:

1. It permits exploitation of superior female

genotype giving more offspring from the same genetic donor than would arise under normal breeding condition.
2. Twin production, introduction of new gene

into cloned herds, manipulation of embryo and transgenic animals have been possible by EIT.
3. Infertility, especially of the older animals

is a major handicap in animal breeding 4/28/12 programme. Such animals of superior

6. It increases the reproductive capacity

of a superior heifer or a cow, since semen from one male can be used for a large number of females. A single superior female can be made to give a large number of ovules through superovulation.
7. Old superior cow that are unable to

maintain pregnancy can still donate ovules for embryo transfer.

8. We may not need to import a superior

quality cow, instead we may have imported frozen embryo which can grow 4/28/12

9. In an animal (cow and buffalo) in vitro

production of embryo has considerable potential value in for disseminating genetic improvement and shortening the generation interval as compared to progeny testing.
10. ETT can be used to resolve several

reproductive enigmas like embryo-utero relationship, endocrine requirement for maintenance of pregnancy, the relative influence of genetic and environment on fetal growth and the biology of zone 4/28/12 pellucida and blastomere.

http:// http://

www.disability-resource.com/medical-hea www.bocafertility.com/gonadotropin.aspx
http://

men.webmd.com/testicular-biopsy

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