WELCOME1

Status of major diseases

(NIH, United States, 2013)2

Heart diseases

Diabetes Parkinson's Cancer AIDS Other0

10

20

30

40

50

60

70

80

Num

ber o

f pati

ents

(in

mill

ions

)

Advances in stem cell technology

Manglam Arya2014-11-102

Centre for Plant Biotechnology and Molecular BiologyCollege of Horticulture

3

Outline 1. Introduction to stem cells2. Types of stem cells 3. Types of human stem cells4. Induced pluripotent stem cells (iPSCs)5. Potential uses of iPSCs6. Success stories 7. Stem cell banking8. Plant stem cell technology and its application9. Summary 10. Conclusion

4

Introduction to stem cells

Term given by William Sedgwick in 1886

Undifferentiated autologous cells

Self renewable capacity

Differentiated • Cells • Tissue • Organ

Show plasticity

5

Types of stem cells Based on their ability

6

Types of human stem cellsEmbryonic stem cells (ESCs) Adult stem cells (ASCs)

Strengths Pluripotent High replicative capacity

Weakness Immunological concerns Subjected to ethical debate

Strengths Multipotent and autologous Clinical safety

Weakness Limited number Limited replicative capacity

7(Diecke et al., 2014)

Induced pluripotent stem cells (iPSCs)

Recently developed stem cells

Genetic modification of adult somatic

cells

Similar as embryonic stem cells

Differentiated into 3 germ layers

Noncontroversial

(Mariusz et al. 2014) 8

Global publications on stem cells

(Servellen and Oba, 2014)9

2005 2006 2007 2008 2009 2010 2011 20120

200

400

600

800

1000

1200

Human embryonic stem cells (hESc) iPSCs

Year

Num

ber o

f pub

licati

ons

Compound Annual Growth Rate(CAGR) for stem cells publication (2008-2012)

(Servellen and Oba, 2014)10

Human embryonic stem cells (hESc)

12%

iPSCs88%

Potential uses of iPSCs

iPSCs Embryonic stem cells

Others- basic research and disease modeling

(Servellen and Oba, 2014)

11

Regenerative medicine

"Process of replacing, engineering or regenerating human cells,

tissues or organs to restore or establish normal function”

– iPSCs are mostly used

– Tissues and organs grown in the laboratory

– Solves the problem of organ shortage

12

(Diecke et al., 2014)

…regenerative medicine

Regenerative medicine

Cardiac disease

Diabetes

Lung disease

Parkinson’sLiver disease

Spinal cord injury

Cancer

13(Ramachandran and Yellededahalli, 2011)

Treatment of type -1 diabetes (T-1D)

• Uses human iPSCs

• Treated with activin A

• Develops insulin producing cells (IPC)

• All cells expressed Pdx-1 genes

• Injected into mouse kidney capsules

(Kim et al., 2015)

iPSCs colony Definitive endodermal

Pancreaticendodermal

Pro-endocrine progenitors

Pre-insulin producing cells

Insulin producing cells

14

Result

Control has 400–500 mg/dl blood glucose

IPC transplanted <200 mg/dl in 3-6 mice

iPSCs are novel option for the treatment of T-1D

Bloo

d gl

ucos

e (m

g/dl

)

Time (minutes)

Control

IPC- transplanted

Glucose clearance

(Kim et al., 2015)15

Treatment for AIDS

(Holt et al., 2010)

AIDS virus relies on a protein CCR5 (C-C chemokine receptor-5)

Hematopoitic stem/ progenitor cells (HSPCs)

16

…treatment for AIDS

(Holt et al., 2010)17

Thymus Lung Spleen Bone marrow0

200400600800

100012001400160018002000

Hum

an C

D4+

T- ce

lls (n

o. o

f ce

lls p

er 5

000)

Other developed regenerative medicineTissue or organ

developed Stem

cells usedUses Reference

Liver iPSCs Acute liver failure Chen et al., 2015

Kidney organoid iPSCs Nephrotoxicity screening, disease modeling

Takasato et al., 2015

Dopamine neuron ESC Parkinson’s disease Kriks et al., 2014

Cerebral organoid (model human brain)

iPSCs Disease modeling and treatment of brain diseases

Lancaster et al., 2013

Neurons iPSCs Neural disorder Corti et al., 2012

18

iPSCs derived functional liver Cerebral organoid

Kidney organoid iPSCs derived neurons19

…other developed regenerative medicine

Stem cells based gene therapy

• Genetic engineering technology for treatment of human

diseases dates to 1989

• Gene therapy has side effects

• iPSCs - new upgraded cellular vehicle or vector

• Homing effect

(Kim et al., 2014)

20

Delivering of therapeutic transgenes

(Kim et al., 2014) 21

iPSCs

Drug screening and development

• iPSCs technology revolutionizing medical science

(Jered et al.,

2014)

• Mechanisms and novel therapeutic molecular targets

• Opportunities for drug discovery

• Large chemical libraries to a list of candidate compounds

(Inoue and Yamanaka, 2011)

22

…drug screening and development

(Faye et al., 2014) 23

Success stories

• Red blood cells created from iPSCs

• In Britain- multimillion pound project

(Hough, 2010)

• American company City of Hope

• FAD approval for human trial for Treatment of HIV and diabetes

(Nicole and White, 2015)

24

• Use of viral vector for gene delivery

• Genetic abnormality in the cells

• Formation of tumor-tumorigenesis

• Very low efficiency of reprogramming

25

Disadvantage of iPSCs

(Nagvenkar and Sriraman, 2013)

Stem cell research institutes in India

Name of institute Place

Centre for Stem Cell Science (CSCS) Hyderabad

International Centre for Stem Cell, Cancer and Biotechnology (ICSCCB) Pune

National Centre for Cell Science (NCCS) Pune

Institute for Stem Cell Biology and Regenerative Medicine (inStem) Bengaluru

Centre for Cellular & Molecular Biology (CCMB) Hyderabad

26

Stem cell banking

• Stores stem cells at very low

temperature

• Regenerative medicine- develops

bioeconomy

• Destined as pillar of the bioeconomy

(Diecke et al., 2014)

27

Steps in cord blood banking

Umbillical cord cut Cord blood collected Test are conducted for maximum safety

Processed to yield maximum stem cells

Stored in liquid nitrogen

(Knoppers and Isasi, 2010)

28

1

54

32

Stem cell regulation in India

• Guidelines prepared by DBT and ICMR

• Conduct clinical trials

• License issued by Indian Institute of Science, Bengaluru

29

(Jayaraman, 2014)

Plant stem cell technology

30

Plant stem cells

• Totipotent in nature

• Located in growing apices

• Grouped into niches- meristem

• WUS(WUSCHEL) gene- signal for proliferation

• CLV3(CLAVATA3) gene controls the expression of WUS gene

31

….plant stem cells

Stem cells niches

WUS protein

Quiescent center

Stem cells in apical meristem Stem cells in root meristem

(Sablowski, 2010)32

Applications

• Tissue culture

• Protect the endangered species

Plant stem cells extract

– Longevity of skin stem cells

– Promote skin’s ability to regenerate

– Delay biological ageing

33

• Uttwiler Spatlauber

• Stem cells taken from fruit

• Extract obtained after lysis of cells

• Incorporated with lecithin liposomes

Apple stem cells as anti-ageing ingredient

(Schmid et al., 2008)

34

Fruit of Uttwiler Spatlauber

Control 0.01 0.102468

101214

Effect of Uttwiler Spatlauber stem cells extract on viability of umbilical cord stem cells

Concentration (in per cent)

cell

num

ber (

in la

khs)

Protection of human stem cells Extract of stem cells was tested for cell viability

(Schmid et al., 2008)35

Control 0.01 0.10

102030405060708090

100

Uttwiler Spatlauber stem cells extract protects umbilical blood stem cells from UV- light

Concentration (in per cent)

Decr

ease

in c

ell n

umbe

r (in

per

cent

)…protection of human stem cells

(Schmid et al., 2008)

Effect of stem cells extract on UV-light treated cells

36

Reversion of senescence signs in fibroblast cells

Gene After H2O2 treatmentcontrol

(% to untreated)

After H2O2+ 2% stem cell extract

(% to untreated)Cyclin B1 (induces proliferation)

73 130

Cyclin E1(controls cells cycle)

78 135

Insulin like growth factor II ( cells proliferation enhancer)

71 117

P53 (tumor suppressor gene)

63 137

Heme oxigenase 1 ( antioxident enzyme)

89 211

(Schmid et al., 2008)37

Plant stem cells product in market

38

Origin Trade name Source

Panax ginseng ActostemR Wkgbase ACT, Korea

Cyperus papyrus Crodarom Papyrus Crodarom, France

Centella asiatica Centella asiatica Stems GTMIRB, Italy

Rhododendron sp. Phyto cellsTectTM Alp RoseMibelle Biochemistry, Switzerland

Malus domestica Phyto cellsTecTM Malus

Vitis vinifera Phyto cellsTecTM Solar Vitis

39

Summary

• Stem cells and its types

• iPSCs – newly developed stem cells

• Development of iPSCs and application in regenerative medicine

• Stem cell banking and its benefits

• Plant stem cell technology in cosmetics

40

Conclusion

• Enormous opportunities for clinical applications

• Biological, ethical and financial challenges

• Unblocks the potential of iPSCs

• Common medicine for future

41

THANK YOU

42

STEM CELLS CAN CREATE MIRACLES…

GIVE HOPE TO THE HOPELESS…

&

COULD MAKE LIFE IMMORTAL!!!