Fertility NEWS LETTER

Ideal fertility : ICSI / IVF & Genetic Center India

Dear Colleges
Hello

I wish you all a very happy and prosperous Deepawali.

In this issue we are discussing two topics.

First is defining an Embryo.Stem cell therapy is hot topic now a days . The best cells are from the growing embryos ,but it creates a big ethical and moral debate. Whether we should have any right to destroy an entity which might have created a full individual if given proper environment. Our embryologist ,Dr.Mrs.Rinku Banerji is not ready to experiment with any of the blastocyst as she feels all embryos are individual and are life's. But there is other view too. My neurosurgeon friends wants stem cells to put them at the site of spinal injury of a paraplegic patient. One is correct ethically and other is correct medically.( This may be a wishful thinking from our lab as we are not in a big research in Stem cell technology ) .Hence there should be definition of Embryo .Who is that ? When life begins ? Should we call a embryo a life only when it is produced by sexual reproduction ( union of sperm and oocyte) and not those by parthenogenesis or Somatic cell nuclear transfer (Cloning).But they have also created life by recent techniques. We tried to find out some answers.

Other is one of the important step in IVF procedure, ovum pick-up. Whole embryology depends on good oocytes and it is the clinician’s responsibility to hand over good number of quality oocytes to their embryologist colleagues.

I wish you all the best.

Dr. D’Pankar Banerji

(Click Archives for Earlier issues)

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What is an Embryo - Current perspective

The definition of embryo is very much required because of technical and more importantly for ethical purpose. Embryonic cells are of tremendous potential and can cure various degenerative diseases. This application may start a business by selling the cell lines by various centers involved in human embryo biotechnology.

What is an human embryo? The answer to the question is “normally” straight forward : The entity created by the fertilization of human oocyte by a human sperm, and when it grows ,it makes a new individual.

But because of development of the reproductive technologies ,the entities has been created that can make a new individual but themselves, not created by regular fertilization.

The definition of the human embryo is proposed as:

A human embryo is a discrete entity that has arisen from either:

1. The first mitotic division when fertilization of a human oocyte by a human sperm is complete or
2. Any other process that initiates organized development of a biological entity with a human nuclear genome or altered human nuclear genome that has the potential to develop up to ,or beyond ,the stage at which the primitive streak appears,
3. And has not yet reached 8 weeks of development since the first mitotic division.

In reproductive technology ,the embryos are made mostly by three types.

1. That is fertilization of the egg by a sperm.
2. That is produced by fusion of polar body and the oocyte itself and makes it diploid ,the process is parthenogenesis. This method of reproduction is present in lower animals and
3. That ,Oocyte is enucleated and empty oocyte is fused with any somatic diploid nucleus.the process call Somatic cell nuclear transfer,a sheep,Dolly was created in this way.

As all these may create an individual ,hence there may be ethical , moral and legal questioning of the use of cells of these early embryos for stem cell therapy or for other researches.

To avoid these problems there are emerging technologies that could produce entities that :

1. Have no potential to implant or result in a live birth and /or
2. Do not have a contribution of genetic information from a sperm and oocyte and/or
3. May contain DNA from two different species

Collection of oocytes in In-vitro fertilization

Collection of oocytes is the crucial step in the whole procedure of test tube baby.
Anesthesia : It is done under the effect of Injection Propopfol or Pentathol Patient is placed in lithotomy position.

Egg retrieval ais a relatively easy and safe procedure. It is scheduled as an pitpatient procedure 35 to 36 hours after the hCG injection.

Patients are advised not to eat or drink for at least 5-6 hours before the procedure. Retrieval is done with the aid of transvaginal ultrasonography ,under intravenous sedation with propofol. The vagina is cleansed with gauze soaked in normal saline, because antiseptics are potentially toxic to the eggs. The patient will also receive a dose of intravenous antibiotic for prophylaxis of pelvic infections.

The entire set for the aspirations prepared on a sterile table, and the needle and tubing are connected to an aspiration pump. The aspiration can be carried with either a single or double lumen needle.

At our center we use single lumen needle from Cook Australia. The aspiration can also be done with the help of 20 ml syringe . The collection of oocytes are done in a fluid called oocyte buffer .It is HEPES buffered media .The needle is washed with the media before actual aspiration begins. When there is good speed of aspiration and identification of oocytes ,the aspiration can be done even without media.

The important thing is that the suction pressure should not go beyond 110 mmHg ,to reduce the fracture of the oocytes.

Each follicle is aspirated in a Falcon tube and is handed to the biologist to check for the presence of the oocyte. If no oocytes are found after aspirating a couple of follicles , then it is important to perform follicular flushing . Follicular flushing is also performed when the number of follicles and eggs is limited. In general a retrieval rate of 70 % to 80% is considered adequate.

Occasionally , there are no eggs despite multiple follicular flushing. In this case, it is important to consider the possibility of improper dosage of hCG . The clinician should request an immediate dosage of hCG , and if low or nonexistent , the retrieval should be interrupted and the patient should be rescheduled 35 to 35 hours after a second ( real ) dose of hCG. It is our experience at Ideal Fertility that this usually gives poorer oocytes .Hence hCG dose is crucial for good oocytes.

It is also very crucial that the collected follicular fluid remains in 37 deg cel.during procedure. Transient falls in temperature disturbs the meiotic spindle and hampers the further cell division and increase the chances of aneupoidy.

Training in IVF and Embryology

Module I : Ovulation induction and Intra Uterine Insemination ( One day )
Module II : Conventional IVF and fundamentals of Embryology( Two days )
Module III : Intra cytoplasmic sperm injection, Micro manipulation ( Two days )

Course Objectives:

The purpose of the course will be to provide an avenue for participants to enter into the field of reproductive biology and an opportunity to gain greater appreciation of the biological processes of mammalian reproduction .At the end of the course, the participants should able to do In-vitro Fertilization of mammalian oocyte.

Course Details :

1. Basic reproductive endocrinology of male and female
2. Ovulation induction and controlled ovarian Hyperstimulation .
3. Sperm preparation methods
4. Fundamentals of embryology
5. Meiosis and its genetic aspect
6. Gametogenesis
7. The development of Assisted reproduction technology
8. IVF culture media and techniques
9. Quality control of embryology lab
10. Handling and culture of Oocytes
11. Stereo zoom and inverted microscopy and photomicrography
12. Setting of the micro manipulators.
13. Intra cytoplasmic Sperm Injection
14. Students will have practical hands on experience in this field and supposed to complete their task and submit a project report at the end.
15. Library and internet facilities will be provided as required.

Course fees :

Module I : Rs.2000.00 ( US$ 50 )
Module II : Rs.20,000.00 ( US$ 500 )
Module III : Rs. 50,000.00 ( US$ 1250 )

For Module I and II Rs.20,000 ( US$ 500 )
For all the three modules/Module II and III : Rs.55,000.00 ( US$ 1375 )

Payments :
Draft : in the name of Dr. D’Pankar Banerji,payable at Jabalpur

Accommodation :
Participants can be provided accommodation in nearby hotels at an extra cost ,Range is Rs.700-3000 per day

Lunch will be served without an extra cost

Timing :
10.00 am to 5.00 pm

ONLY TWO PARTICIPANTS PER BATCH FOR MODULE II AND III

Dates :
Throughout the year.

Course Directors :
Dr. D’Pankar Banerji,IVF specialist and Dr.Mrs.Rinku Banerji,Embryologist and molecular pathologist.