IMA

1. STEM CELLS AND INFERTILITY
Dr. Gayathiri Ganesan Ram
MS(O.G), Fellow in Reproductive Medicine and
Andrology.
Consultant Reproductive Medicine Specialist
ARC Fertility Hospitals

2. ‘68
‘78
‘81
‘94
‘95
‘96
‘98
‘01
‘05
‘06
‘07
Edwards and Bavister fertilize
first human egg
Mouse embryonic stem cells
derived
Non-human primate ES cells
derived
Thomson et al derive and
culture first hES cells
Woo-suk Hwang fakes human
stem cell cloning
Yamanaka et al derive human
iPS cells
First IVF baby Louise Brown
is born
First donated blastocysts
for research
Dolly, the cloned sheep, is
born at the Roslin Institute
Mouse embryonic stem cells
derived by SCNT
Yamanaka et al derive
mouse iPS cells
A Walk Through the History…

3. Human embryonic stem cell: Also known as a
human pluripotent stem cell, one of the "cells
that are self-replicating, are derived from human
embryos or human fetal tissue, and are known to
develop into cells and tissues of the three
primary germ layers."
(From the National Institutes of Health Guidelines for Research Using Human Pluripotent Stem Cells.)
Definition

4. Human Embryonic Stem Cells.
• Definition: Stem cells are
clonogenic,undifferentiated, self replicating,
progenitor cells, capable of self regeneration
and also of multilineage differentiation and
generation of several other tissues and organs
in the body

5. Types of Stem Cells.
• 1) Toti potent stem cells.
• 2) Pluri potent stem cells.
• 3) multi potent stem cells.
• 4) Unipotent stem cells- oxymoron?

6. Totipotent Stem Cells.
• Natural identical twinning is due to division of
an early embryo into two embryos.
• One form of cloning would be by dividing an
early embryo into two or more embryos.

7. Toti Potent Stem Cells
• The embryonic cells
in the early stages are
toti potent. Eg: two
celled embryo,four
celled embryo.

8. Pluripotent Stem Cells
• Pluri potent cells are capable of producing all
the cells, tissues and organs in the body- but
not an embryo.
• Cells obtained from the blastocyst -from the
inner cell mass are-pluri potent.

9. Pluri Potent Stem Cells
• The cells from the
inner cell mass are
pluri potent. They are
capable of producing
any organ or tissue in
the body-but not the
placenta.

10. Stem Cell Cultivation
• The slide briefly
describes embryonal
stem cell cultivation.
• University of
Wisconsin was the
first one to cultivate
human embryonic
stem cells.

11. Embryonic Stem Cells

12. Multi Potent Stem Cells
• Adult stem cells, cord blood stem cells,and
cells obtained from late stage embryos are
multi potent.
• Multi potent stem cells are capable of
generation of several tissues, but not all
tissues.

13. Uni potent Stem Cells
• Oxymoron?
• The stem cells found in many organs like
liver,bone marrow,testes serve as reserve cells
for regeneration of appropriate tissue.
• Under proper in vitro/in vivo conditions these
cells can be coaxed to become other tissues of
the body.

14. Embryonic Stem Cells
• These cells are obtained usually from the
inner cell mass of a blastocyst- day 6/7
embryo.
• These embryos are usually spare embryos,
research embryos or orphaned
embryos,obtained from an infertility clinic.

15. Sources of Stem Cells
• 1)Embryonal stem cells.
• 2)Adult stem cells.
• 3)Cord blood stem cells.
• 4)Induced pluripotent stem cells
• 5)Induced multipotent stem cells
(transdifferentiation)
• 6)Somatic cell nuclear transfer (Dolly)
• 7)Oocyte, somatic cell fusion -
http://news.sciencemag.org/sciencenow/2011/10/a
n-egg-citing-recipe-for-human-s.html

16. Adult Stem Cells
• These cells are generally obtained from bone
marrow.They may also be obtained from
peripheral blood.
• Stem cells are probably present in most of the
tissues and organs in the body.
• It is difficult to identify, isolate culture and
grow adult stem cells.

17. Adult Stem Cells
• Identifying and isolating adult stem cells from
many tissue would be worse than the
proverbial search for ‘needle in a haystack.’
• If this becomes possible, this would avoid
many of the legal, ethical and moral problems
associated with the use of Embryonal stem
cells.

18. Adult Stem Cells
http://stemcells.nih.gov/

19. Cord Blood Stem Cells
• Cord blood is a good source of stem cells.
• It may be a good idea to store cord blood
routinely at all deliveries for autologous stem
cell transplant or HLA compatible stem cell
transplant- where the older sibling/ other
family members are afflicted with
haematological abnormalities/ malignancy.

20. Indications for Stem Cell Therapy.
• Stem cell therapy can be used in many
degenerative diseases.
• Stem cells can be used in the regeneration of
many diseased organs.
• Stem cells can be used in the correction of
many metabolic and haematologic disorders.

21. Indications for Stem Cell Therapy.
• Current:
• 1)diabetes mellitus- insulin dependent.
• 2)Parkinson’s disease.
• 3)heart disease- both ischaemic and
degenerative.

22. Indications for Stem Cell
Therapy.
• 4)spinal cord injuries.
• 5)Haematologic disorders- like Fanconi’s
anaemia,Thalassaemia.
• 6)malignancy.
• 7)autoimmune diseases.
• 8)genetic diseases.

23. Indications for Stem Cell
Therapy.
• Virtually any diseased organ or tissue can be
replaced with stem cell therapy, obviating the
need for transplants.
• Future research holds very great promise for
correcting many diseases.
• A new specialty is emerging “regenerative
medicine’ thanks to the advent of stem cell
therapy.

24. Mode of Administration of Stem Cells.
• 1)direct injection into the organ.
• 2)systemic administration and the stem cells
colonize the organ/tissue.
• 3)direct surgical placement of stem cells.

25. Somatic Cell Nuclear Transfer

26. • Allows for avoidance of immune problems
• Provides patient-specific disease models for
research
• Could lead to reproductive cloning
• Needs the donation of eggs
SCNT

27. Stem cells and the reproductive tract
• Stem cells and the endometrium
• In humans, regeneration of the endometrium
occurs monthly during reproductive life and
beyond under appropriate ovarian steroid
priming.
• Cyclic replenishment of the cellular
compartments of the endometrial functionality
by adult stem cells (ASC) is essential for the
preparation of this organ for its main function,
i.e. allowing implantation and pregnancy to
proceed

28. • ASC present a MSC phenotype (Chan et al.,
2004; Schwab and Gargett, 2007; Schwab et
al., 2008; Cervello et al., 2010)
• They functionally contribute to human
endometrial regeneration in vitro and in vivo
(Cervello et al., 2010, 2011; Matsuda and Shi,
2010).

29. • Bone marrow-derived stem cells (BMDSCs) have been
shown to contribute as an exogenous source to tissue
repair and regeneration of different organs and tissues
(Pittenger et al., 1999).
• In the human, BMDSCs are also a source of non-
hematopoietic cells in the different endometrial cellular
compartments (stroma, glandular epithelium, and luminal
epithelium).
• They contribute mainly to the formation of endometrial
stromal compartment cells and to a much lesser extent to
the glandular and luminal epithelium (Du and Taylor, 2007;
Mints et al.,2008; Ikoma et al., 2009; Cervello et al., 2012).

30. • It has been recently proposed that BMDSC
infusion might improve endometrial regeneration
in a murine model of AS (Alawadhi et al., 2014;
Jing et al., 2014; Kilic et al.,2014; Zhao et al.,
2015).
• However, peripheral blood stem cell
transplant(PBSCT) did not result in engraftment
of donor stemcells in the recipient uterus in a
macaque model, or human (Wolff et al., 2013).

31. SPERMATOZOA
• Obtaining PGCs (primodial germ cells) from pluripotent
cells is the first step in the differentiation toward post-
meiotic spermatozoa. However, further differentiation
has proven challenging in vitro.
• The strategies employed to favor it include
supplementing the differentiation medium with the
growth factors bone morphogenetic protein (BMP)-4, -
7, -8b (Geijsen et al.,2004; Tilgner et al., 2008), N2B27,
activin, Fibroblast growth factor 2 (FGF2) (Hayashi et
al., 2011), retinoid acid (Nayernia et al., 2006;Eguizabal
et al., 2009, 2011; Cai et al., 2013; Peng et al.,
2013),R115866 (Eguizabal et al., 2011), and hormones
such as Insulin andTestosterone (Easley et al., 2012).

32. STEM CELLS AND VAGINAL
RECONSTRUCTION
• The use of stem cells for vaginal recovery has
been poorly investigated so far, but most work
has been carried out with muscle-derived stem
cells (MDSC).
• Recently, muscle has been identified as an
alternative source of adult stem cells, different
from satellite cells, possessing the capacity to
differentiate into different cell lineages.
• MDSCs appear as promising candidates for the
treatment of muscular, cardiac and urological
disorders.

33. STEM CELLS AND ERECTILE
DYSFUNCTION
• Penile erection is a neurovascular response
involving an increase in arterial inflow, relaxation
of corpora smooth muscles, and a reduction
invenous outflow.
• Erectile dysfunction can be caused by conditions
such as diabetes, neuropathies, cavernous nerve
injuries, neurotransmitterimbalances, vascular
diseases, inflammation diseases (Peyronie
• disease) or surgery.

34. • The results obtained indicated that the stem cell
injections brought benefits in terms of restored
erectile function and penile physiology (Zhang et
al., 2012); interestingly, the improvement in
erectile function seems to be due to the
paracrine factors secreted by the injected cells
(cytoprotective, anti-fibrotic and anti-apoptotic
molecules),rather than direct
grafting/differentiation (Albersen et al., 2010).
• No informationis available on the duration of the
beneficial effects.

35. • The only preclinical study involving patients was
carried out with intracavernosal injections of
umbilical cord blood stem cells in seven patients
with diabetic-erectile dysfunction: patients
reported an improvement in penile erection and
an increase in penile rigidity, especially when in
combination with PDE5 inhibitors; the duration of
the effect was variable but after a few months all
patients experienced a regression
• (Bahk et al.,2010).

36. Current Research
• 1) current research is aimed at identifying,
isolating, culturing and growing embryonal
stem cells and adult stem cells.
• 2) identifying organizers/ growth factors which
direct stem cells to grow into different tissue
and organs.

37. Current Research
• 3)modifying adult multi potent stem cells to pluri
potent stem cells.
• 4) engineering stem cells to avoid expression of HLA
antigen which often lead to immune rejection.
• 5) optimizing the conditions for inducing
pluripotyency by expressing transgenes in somatic
cells
• 6) carefully characterizing the genetic and epigenetic
abnormalities of current stem cells to avoid
malignancy in the patient

38.  2002
◦ Stem Cells "Cure" Diabetes in Mouse – Stanford
◦ Stem Cells Improve Motor Function in Rat Model of
Parkinson's Disease - NIH
 2003
◦ Mouse Embryonic Stem Cells Develop Into Sperm – Japan
◦ Human Embryonic Germ Cells Restore Movement to
Paralyzed Rats – Johns Hopkins
 2004
◦ Heart Muscle Cells Produced from Human Embryonic Stem
Cells Can Replace Biological Pacemaker – Israel
◦ Mouse Sperm Generated from Stem Cells Able to Fertilize
Mouse Egg - Harvard
Research Highlights of the Field

39.  2005
◦ Embryonic Stem Cells Cure Mouse Model of Hemophilia – UNC
Chapel Hill
◦ Alternative Method for Deriving Stem Cells Proven in Mice – MIT
 2006
◦ Scientists Generate Human Embryonic Stem Cell Lines from
Single Cells – Advance Cell Technology
◦ Scientists Reprogram Adult Mouse Skin Cells by Adding Defined
Factors – Japan
 2007
◦ Reprogrammed Mouse Skin Cells Cure Mouse Sickle Cell Anemia
– MIT
◦ Human Skin Cells Reprogrammed – Japan
Research Highlights of the Field

40. • 2008
– Scientists Generate Stem Cell Line from Patient
with ALS Disease – Harvard
– Adult Stem Cell Lines Created for 10 Additional
Human Diseases – Harvard
• 2009
– Geron Receives FDA Clearance to Begin World's
First Human Clinical Trial of Embryonic Stem Cell-
Based Therapy
Research Highlights of the Field