The EBAI CME 2013 - 21st and 22nd of September 2013, Golden Valley Resort, Ghodbunder Road, Thane West, Mumbai.

1. Role Of Eye Bank Beyond TheRole Of Eye Bank Beyond The
Cornea-STEM CELLSCornea-STEM CELLS
Dr.Srivalli Kaza
21st
September 2013
National EBAI CME, Thane.

2. ACKNOWLEDGEMENTSACKNOWLEDGEMENTS
WAVIKAR EYE INSTITUTE,
THANE
L.V.PRASAD EYE INSTITUTE,
HYDERABAD

3. Dr.Virender S SangwanDr.Virender S Sangwan
Shanti Swaroop
Bhatnagar Award 2006
Biotech Products and
Process Development
and Commercialization
Award 2007
ACKNOWLEDGEMENTSACKNOWLEDGEMENTS

4. What Are Stem Cells?What Are Stem Cells?
Stem cells are the raw
material from which all of
the body’s mature,
differentiated cells are
made.
Stem cells give rise to
brain cells, nerve cells,
heart cells, pancreatic
cells, etc.

5. Stem Cell – DefinitionStem Cell – Definition
A cell that has the
ability to
continuously divide
and differentiate
(develop) into
various other kind(s)
of cells/tissues

6. Stem Cell – are DynamicStem Cell – are Dynamic
Are undifferentiated
“master” cell that do not
yet have a specific
function
Can change to one or
several different cell
types (differentiate)
under proper conditions
Can undergo unlimited cell
division, self-renewal)
Stem cell
Stem cell
Self-renewal
Specialized cell
(e.g., white blood cell)
Differentiate

7. Major types of Stem CellsMajor types of Stem Cells
The two broad types of
mammalian stem cells are:
(a)embryonic stem cells that
are isolated from the inner
cell mass of blastocysts,
(b)adult stem cells that are
found in adult tissues. In a
developing embryo, stem
cells can differentiate into
all of the specialized
embryonic tissues. I

8. Autologous – Stem CellsAutologous – Stem Cells
Sources of the patient's own
stem cells (autologous) are
either the cells from
patient's own body or his
or her cord blood. For
autologous transplants
physicians now usually
collect stem cells from the
peripheral blood rather
than the marrow
This procedure is easier,
unlike a bone marrow
harvest, it can take place
outside of an operating
room and the patient does
not have to be under
general anaesthesia.

9. Allogeneic – Stem CellsAllogeneic – Stem Cells
 Sources of stem cells from
another donor (allogeneic)
are primarily relatives
(familial-allogeneic) or
completely unrelated
donors (unrelated-
allogeneic). The stem cells
in this situation are
extracted from either the
donor's body or cord blood

10. Xenogenic - Stem CellsXenogenic - Stem Cells
In this stem cells from
different species are
transplanted, e.g. striatal
porcine fetal ventral
mesencephalic (FVM)
xenotransplants for
Parkinson's disease.
This has no major ethical
concerns and a large
amount of tissue is
available, however life long
immunosupression and risk
of rejection are the major
limitations

11. History of Adult Stem Cell ResearchHistory of Adult Stem Cell Research
Since the 1970’s, bone marrow
transplants have been used for
treatment of Immunodeficient
and leukemia.

12. History of Human EmbryonicHistory of Human Embryonic
Stem Cell ResearchStem Cell Research
In 1998, James Thomson (University of
Wisconsin-Madison) isolated cells from the inner
cell mass of the blastocyst, and developed the first
human embryonic stem cell line in culture.
Isolate inner cell mass
Culture cells
Inner cell mass
(forms fetus)
Day 5-6
Blastocyst

13. What Diseases Can beWhat Diseases Can be
Cured by Stem Cell TherapiesCured by Stem Cell Therapies
Any disease in which
there is tissue
degeneration can be a
potential candidate for
stem cell therapies

14. Major Progress in Several ImportantMajor Progress in Several Important
Health problemsHealth problems
Alzheimer’s disease
Parkinson’s disease
Spinal cord injury
Heart disease
Severe burns
Diabetes

15. Drug TestingDrug Testing
Stem cells could allow scientists to
test new drugs using human cell
line which could speed up new
drug development.
Only drugs that were safe and had
beneficial effects in cell line testing
would graduate to whole animal or
human testing.
It would allow quicker and safer
development of new drugs.

16. Source of Stem Cells for MedicalSource of Stem Cells for Medical
therapiestherapies
Tens of thousands of frozen
embryos are routinely
destroyed when couples
finish their treatment.
These surplus embryos can
be used to produce stem
cells.
Regenerative medical
research aims to develop
these cells into new,
healthy tissue to heal
severe illnesses.

17. Stem Cell Research WorldwideStem Cell Research Worldwide

18. AdultAdult multipotentmultipotent stem cellsstem cells

19. Adult Stem CellsAdult Stem Cells

20. Limbal stem Cell therapyLimbal stem Cell therapy
The treatment is known as
limbal stem cell therapy,
and the patients who
received the treatment
suffered from chemical
burn or genetic disease
know as aniridia
By replacing the limbal stem
cells, the cornea begins to
clear up as the cells are
replaced with the healthy
transparent layer again.
CLAuT – 1 month post-op

22. Limbal stem cellLimbal stem cell
CONJUNCTIVA
CORNEOSCLERAL
LIMBUS
CORNEA
Peripheral Central
TDC
PMC
Differentiable Compartment
Suprabasal cells
®Goblet cells
Proliferative
Compartment:
Basal cells
TAC
SC
TDC
PMC

25. Petri dish
Amniotic membrane
Limbal Tissue
Stem cells
Culture techniqueCulture technique
Feeder
cells

28. AMG & Limbal Stem CellsAMG & Limbal Stem Cells
• More efficient ocular surface
reconstruction in severe
chemical injuries
Ucakhan et al, Cornea 2002

29. LSCT - IndicationsLSCT - Indications
• Chemical Injuries
• Stevens Johnson Syndrome
• Ocular cicatricial
Pemphigoid
• Thermal injuries
• Severe Shield ulcer
• Neurotrophic keratitis
Sridhar et al, Am J Ophthal 2000
Shimmura et al, Cornea 2001
Sridhar et al, Ophthalmolgy 2001
Sippel et al, Curr Opin Ophthalmol 2001

30. Surgical Management-EvolutionSurgical Management-Evolution
 Conjunctival transplantation(Thoft,1977)
 Keratoepithelioplasty(Thoft,1984)
 Limbal Autograft(Kenyon & Tseng,1989)
 Keratolimbal Allograft(Tsai & Tseng,1994)
 Living related conjunctival Allograft (Kwitko et
al 1995)
 Human AMG(Tsubota et al,1996)
 Limbal Stem cell culture (Pellegrini et al, 1997)
 SLET (Sangwan et al, 2012)

31. Case-I

32. 16/10/2001
• 25 Years , Male
• C/O severe photophobia, decreased vision,
watering & burning sensation (OU)-
following injury to his face with Sodium
hydroxide-1& 1/2 Months ago
• Was on Rx with Ciplox e/d, Betagan e/d,
Atropine e/d, Moisol e/d, Predforte e/d,
Norflox e/o & Tab. Vitamin C

33. OD

34. OS

35. Diagnosis
• OU-Chemical Injury(Grade IV)
Persistent Epithelial defect

36. Management
• OU-AMG under LA---16/10/2001
• OD-Limbal Transplantation from living related
donor(Brother)---
04/07/2002(9 & 1/2 months)
• Advice
OD-Prednisolone acetate 1% e/d 1 hourly
Ciprofloxacin hydrochloride 0.3% e/d qid
Tears Plus e/d qid
Tab. Cyclosporin 100 mg 2-2-1
Inj. Methyl Prednisolone 500mg stat

37. OD
OS

38. 05/09/2002
(1 year)
20/125—>20/80
OD-Penetrating Keratoplasty

39. CASE-2

40. 24/09/2001
• OS-Chemical burns(grade II )
Symblepharon
Epithelial Defect
Retained FB
25/09/2001
• OS-Removal of foreign bodies + AMG
under LA

41. 28/02/2002
(5 months)

42. OS

43. PLAN
• OS-Symblepharon release + AMG
under GA on 28/02/2002

44. 30/04/2002
(7 months)
CF @ 1m
OS

45. 23/05/02(8 months)
• OD--Limbal Biopsy under LA
06/06/02(9 months)
• OS--Limbal transplantation under
LA

46. 03/07/2002(11 months)
OS 20/400-->20/100

47. 09/10/2002( 1 year)
OS 20/40-->20/25P

49. Research on Stem Cells isResearch on Stem Cells is progressingprogressing inin
spite of several restrictions!spite of several restrictions!