Chronology of IVF-ICSI Research in INDIA (2022)

Title: Chronology of IVF-ICSI Research in INDIA

In-Vitro Cell Technology in India Origin,
Evolution and New frontiers
Dr. Sudarsan Ghosh Dastidar, MD GD Institute for
Fertility Research Kolkata, India
HEAL Conference, India International Centre , New
Delhi, 23rd September 2009
Assisted Reproductive Technology (ART) - till now

  • Manipulation of human cells (egg and sperm) out
    side the body, i.e., In-Vitro, started with the
    breakthrough of test tube baby technology in
  • Two main modalities
  • IVF
  • ICSI
  • The advanced laboratory set up and technology
    required for this may be the basis for embryonic
    stem cell culture and future cell based therapy.

Chronology of IVF-ICSI Research in INDIA

  • 1978 Dr. Subhas Mukherjee his team.
    Birth of Durga.
  • 1982-1986 Kolkata-Dr. B. N. Chakravorty
  • Dr. S. Ghosh Dastidar
  • Birth of Imran.
  • 1983-1986 Bombay- Dr. I. Hinduja
  • Dr. Anandkumar
  • Birth of Harsha ( July 1986 )
  • 1992 Kolkata- 1st GIFT Baby
  • Dr. S. Ghosh Dastidar Dr. K
    Ghosh Dastidar
  • 1995 Kolkata- 1st ICSI ZIFT Baby
  • Dr. S. Ghosh Dastidar Dr. K
    Ghosh Dastidar
  • 1995 Mumbai- ICSI Baby
  • Dr. Firuza Parikh

Dr. Subhas Mukherjee The pioneer of IVF Research
in India.Responsible for inducting me into the
emerging field of In-Vitro Fertilization and
related science.
World Congress on IVF, Helsinki, Finland, 1984
World Congress on IVF, Istanbul, 2005
Cornell Medical Center, USA Dr. Bedford and
myself, 1987
Prof. Struart Campell, myself and Dr. Kakali
Ghosh Dastidar
Our Initial Struggle (1982)

  • The story of how we developed IVF in India
    (Kolkata) is one of Herculean struggle. It is
    necessary to narrate many personal details which
    are vital to illustrate how breakthroughs in
    creative science really happen, specially in a
    country like ours.
  • Our challenges
  • Non existent infrastructure for IVF related cell
    culture in India. Hence I had to prepare
    tissue culture media from scratch, using the bare
    minimum resources available in Kolkata
  • Frequent power cuts in 1981-82 standby generator
    was must
  • There was no computer and obviously no internet
    thus there was almost no literature/publication/jo
    urnals available in Kolkata
  • The most challenging job was to develop a tissue
    culture system which would produce 5 Co2 in air
    and 95 relative humidity atmosphere since we did
    not have the modern computerized Co2 incubator.
  • Though it may sound extraordinary to researchers
    today, one basic challenge was to first identify
    how the human egg looked under the microscope,
    in any book/journal/internet

Learning by Trial Error

  • During 81-82 we had no knowledge even about
    identification of human egg or the associated
    laboratory procedures for IVF. Thus we had to
    start from scratch.
  • I was entrusted with the responsibility to
    develop basic IVF laboratory aspects like
    Preparation of tissue culture media, Oocyte
    identification sperm preparation
    fertilization embryo-culture.
  • The only basic knowledge I possessed was of a
    theoretical nature, and came from my discussions
    with Late Dr. Subhas Mukherjee.


  • Prof. B. N. Chakraborty was involved in
    developing the clinical aspects like patient
    selection, ovarian stimulation and most
    importantly, oocyte retrieval and finally embryo
    transfer (ET).
  • I was entrusted to develop the embryo culture
    laboratory and then the laboratory methods. My
    major challenges were
  • To identify human oocyte from follicular fluid
  • To device some system to produce 5 CO2 in air
    atmosphere for IVF and embryo culture since we
    did not have modern CO2 incubator.
  • To capacitate spermatozoa an absolute necessary
    step to achieve successful fertilization of
  • To prepare tissue culture media preparation PH
    and osmolarity control of media. Prof. Subir
    Dutta, an eminent pathologist of Kolkata extended
    logistic and technical support.
  • To learn manipulation of human oocyte under
    stereo microscope.
  • Thus, I was a self trained IVF laboratory expert!!

A typical day

  • Morning 5 6 a.m. cleaning of laboratory,
    final preparation of media and other lab
  • 7-9 a.m. basic quality control, preparation for
    egg recovery.
  • 1030 a.m. to late evening - I used to carry the
    test tube containing follicular fluid in a
    thermos flask to a laboratory in far away
    Jadavpur University and later, to Indian
    Statistical Institute, where I was desperately
    trying to identify the oocytes under stereo
    microscope (no one in Calcutta then even knew
    their appearance !)
  • However, soon we realized that in order to
    achieve pregnancy by IVF we needed a laboratory
    close to the clinic where oocyte retrieval was
    being done. Otherwise the temperature and pH of
    the egg-containing follicular fluid was being
    altered in this long transit.
  • Thus, I set up Calcuttas first IVF laboratory
    (very primitive though!) in my study room at
    79/28, A.J.C. Bose Road, Kolkata - 14.
  • Some days I used to finish work at 12 night or
    even later.

Humble Beginnings
and, the modern IVF lab !
The improvised IVF embryo culture system devised
by me in 1983

  • A pre-requisite for IVF is perfectly controlled
    environment for cell culture, currently provided
    by state-of-art incubators as shown in the
    previous slide.
  • During 1982-86 we did not have modern incubator -
    an absolutely essential equipment for human IVF.
  • Thus I tried to improvise several alternative
    simple system with locally available materials.
  • Finally I was able to develop a system to
    produce 5 CO2 in air atmosphere and
    approximately 85 relative humidity.

Improvised embryo culture system developed by me
using 5 CO2 in air environment (1982)
Modern CO2 Incubators used in our center
5 CO2 in air
Test tube containing egg in culture media
Water in petri dish
Initial breakthrough in IVF (1982-1984)

  • After many failures, I achieved fertilization of
    oocyte and its subsequent cleavage to 6-8 cell
    stage in June/July 83.
  • Following embryo transfer, a pregnancy occurred.
    (September 83). Unfortunately, this pregnancy
    ended in abortion at around 10 weeks.
  • It was a major break through indicating the
    feasibility of IVF as a clinical method to treat
    tubal factor infertility.
  • Dr. B. N. Chakraborty and myself jointly
    presented this data in 3rd World Congress on
    IVF, Helsinki, Finland, May 1984.
  • This was the 1st report of successful IVF and
    embryo transfer resulting in pregnancy in human
    from India in any World Congress. (May84)
  • Ref. Our Experience of IVF in India Chakraborty
    B.N. and Ghosh Dastidar S. Congress book 3rd
    World Congress on IVF, Helsinki, Finland, May 1984

Microphotographic Documents of oocytes taken by
me during 1982-1983
Offshoot of IVF Technology novel sperm
preparation and IUI in human infertility

  • The technology of sperm preparation in IVF,
    prompted me to venture introducing 100 motile
    sperm in uterine cavity in patients with
    unexplained infertility.
  • I was surprised to achieve series of pregnancies
    following such method during 1982-84.
  • These were the very first few IUI
    pregnancies in the world.

Abstract of our paper presented at 5th World
Congress on Human Reproduction Athens, Greece,
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ICSI the major breakthrough in Assisted
Reproductive Technology after IVF

  • Till 1993 there was no treatment for severe male
    factor infertility with very low sperm
    count/motility, not even by IVF
  • This changed with the advent of Intra Cytoplasmic
    Sperm Injection (ICSI) which means fertilizing an
    egg by micro injection with a single viable
    spermatozoa in the laboratory, first reported in
    1992-93 from Brussels, Belgium.
  • I initiated one of Indias first two ICSI
    programs in Kolkata in January 1994, which was
    responsible for the birth of a ICSI-ZIFT baby in
    March 1995.

Intra Cytoplasmic Sperm Injection
Establishment of Successful ICSI in Our Center,
Our first ICSI baby in 1995
Different stages of fertilized egg and embryo
following IVF/ICSI documented in my Lab
4 cell stage
PN Stage
8 cell stage
What next ?
Blastocyst embryo
Morula stage
Blastocyst stage
Human Embryonic Stem Cell (hESC)

  • Stem cells are unspecialized.One of the
    fundamental properties of a stem cell is that it
    does not have any tissue-specific structures that
    allow it to perform specialized functions. For
    example, a stem cell cannot carry oxygen
    molecules through the bloodstream (like a red
    blood cell).
  • Stem cells can give rise to specialized cells.
    When unspecialized stem cells give rise to
    specialized cells, the process is
  • Scientists are just beginning to understand the
    signals inside and outside cells that trigger
    each step of the differentiation process.
  • The internalsignalsare controlled by a
    cell'sgenes, which are interspersed across long
    strands of DNA, and carry coded instructions for
    all cellular structures and functions.

Historical Landmarks
Scientists discovered ways to derive embryonic
stem cells from early mouse embryos
The detailed study of the biology of mouse stem
cells led to the discovery of a method to derive
stem cells from human embryos and grow the cells
in the laboratory. These cells are calledhuman
embryonic stem cells.
Major breakthrough - identification of
conditions that allow some specialized adult
cells to be "reprogrammed" genetically to assume
a stem cell-like state. This new type of stem
cell, calledinduced pluripotent stem cells
How are embryonic stem cells grown in the
Growing cells in the laboratory is known ascell
culture. Human embryonic stem cells are isolated
by transferring theinner cell massfrom the
blastocyst into a plastic laboratory culture dish
that contains a nutrient broth known asculture
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Pluripotency the God of small cells

  • Human Embryonic stem cells, as their name
    suggests, are derived from 4-5 days old surplus
    human embryos produced by IVF centers.
  • They self-renew indefinitely in the
    undifferentiated state producing millions of
  • They are pluripotent, which means they can
    develop into all cells and tissues in the body.
    Have unique regenerative abilities
  • The most important potential application Cell
    Based Therapies in
  • 1) Diabetes
  • 2) Alzheimers disease
  • 3) Spinal cord injury
  • 4) Stroke
  • 5) Heart disease
  • 6) Osteoarthritis
  • With further research it may be possible to
    understand how cell proliferation is regulated
    during normal embryonic development or during
    abnormal cell division and differentiation that
    leads to cancer.

The invisible Hands of God the miracle solution
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How are embryonic stem cells stimulated to
Directed differentiation of mouse embryonic stem
Diseases that might be treated by transplanting
cells generated from human embryonic stem cells
includeParkinson's disease, diabetes, traumatic
spinal cord injury,Duchenne's muscular dystrophy,
heart disease, and vision and hearing loss.
What are the potential uses of human embryonic
stem cells and the obstacles that must be
overcome before these potential uses will be

  • Studies ofhuman embryonic stem cellswill yield
    information about the complex events that occur
    during human development.
  • Primary goal - to identify how undifferentiatedst
    em cells become the differentiated cells that
    form the tissues and organs. Scientists know
    that turninggeneson and off is central to this
  • Serious medical conditions, such as cancer and
    birth defects, are due to abnormalcell
  • A more complete understanding of the genetic and
    molecular controls of these processes may yield
    information about how such diseases arise and
    suggest new strategies for therapy.
  • Testing of drugs - new medications could be
    tested for safety on differentiated cells
    generated from humanpluripotentcell lines.

Have human embryonic stem cells successfully
treated any human diseases?

  • January 23, 2009 - FDA granted clearance for
    Gerons (Califormia based pharma company)
    clinical trial of GRNOPC1 in patients with acute
    Spinal Cord injury the Worlds 1st Human
    clinical trial of Embryonic Stem Cell based
  • Gerons 2nd HESC product GRNCM1 involves pre
    clinical trial of cardiomyocytes for treating
    myocardial disease.

In-Vitro Cell Technology in India Origin,
Evolution and New frontiers
Embryonic Stem Cell from Blastocyst
Cell based therapies
Future regenerative medicine

  • Our human body as the supreme model of omni
    potent mechanism.
  • It appears that in near future practice of
    medicine will be rewritten with inclusion of
    regenerative or restorative medicine.
  • The primitive immortal cells that is embryonic
    stem cells appears to have the potential to
    restore the body mechanism from disease process
    by an unique regenerative approach hitherto
    unknown to mankind.

enormous energy
Single atom-------------
?Atom Bomb
Regenerative medicine
Millions or trillions of hESC
Embryonic Stem Cell
? Ethical issues
Research Ethics and Stem Cells

  • The ethical question which arises is to what
    extent should researchers manipulate the most
    basic processes of human life.
  • However, we need to ask ourselves whether the
    potential benefits for treatment of disease and
    improving quality of life outweigh such ethical
  • Thus, it would seem that research in embryonic
    stem cell promises much, and also needs
  • Here we should keep in mind that during early
    days of evolution of IVF technology in India, it
    was precisely the lack of constraining regulation
    which permitted such exponential growth.

Thank you

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