Lineage Restriction in Stem Cells

1
Lineage Restriction in Stem Cells
Principal Investigator Dr Lim Sai Kiang
RESEARCH FOCUS Our research program aims to
generate therapeutically important cell lines
from embryonic stem cells (ESCs) for the
development of cell-based or biologic-based
therapy. This approach circumvents the risk of
ESC-associated teratoma formation and provides
scalable sources of homogenous cell preparations.
Our strategy for generating these lines is
through incremental lineage restriction of ESC by
differentiating and establishing progenitor cell
lines first from ESC and then sequentially from
ESC-derived progenitor lines. Using this
approach, we have successfully isolated
endothelial progenitor cell lines and
insulin-producing cell lines from mouse ESCs, and
mesenchymal stem cell lines from human ESCs. We
are testing these cell lines and their derivative
products in mouse models of acute myocardial
infarction and type 1 diabetes, rabbit models of
osteochondral defects and porcine model of acute
myocardial infarction.
Derivation of insulin-producing cell lines from
mouse ESC
E-RoSH lines
3.5 dpc
Insulin-producing E-RoSHK cell lines
Differentiation
ESCs
Differentiation
EBs
Insulin C-peptide immunostaining of ERoSHK
cells
Lian et al, 2006 PLoS ONE 1e6
Endothelial cells
CD9hi, SSEA-1-
RoSH cell lines
Insulin-producing RoSH2K cell lines
Differentiation
Differentiation
Reversal of hyperglycemia in diabetic mouse by
RoSH2K cell transplantation
Yin et al, 2004 ATVB 24 691- 696.
Endothelial cells
Insulin-producing MEPI cell lines
Differentiation
5.5 6.5 dpc
Glucose-sensing components, glucokinase (GK)
glucose transporter (Glut2) in MEPI cells
Derivation of human mesenchymal stem cell lines
Hues9.E1
FGF2/PDGF
Typical MSC surface antigen profile
trypsinize
Von Kossa
Alcian blue
Oil red
Differentiation
Collagen (II)
MSC
Lian et al, (2007). Stem Cells 25(2)425-36.
ESC
Chondrocytes
Adipocytes
Osteocytes
Conditioned medium (CM)
Reducing reperfusion injury in pig model of AMI
Secretory proteome
?
infarct
CM
Saline
NCM
?
?
?

Oxidative stress
8OHdG
Sze et al, (2007). Molecular and Cellular
Proteomics 61680-1689
Publications (2006 2007) 1. Lian Q, Yeo KS, Que
J, Tan EKY, Yu F, Yin Y, Salto-Tellez M, El
Oakley RM, Lim S-K (2006). Establishing clonal
cell lines with endothelial-like potential from
CD9hi, SSEA-1- cells in embryonic stem
cell-derived embryoid bodies. PLoS ONE 1e6. 2.
Lian Q, Lye E, Yeo KS, Tan EKY, Salto-Tellez M,
Liu TM, Palanisamy N, El Oakley RM, Lee EH, Lim
B, Lim S-K (2007). Derivation of Clinically
Compliant MSCs from CD105, CD24- Differentiated
Human ESCs. Stem Cells 25(2)425-36. 3. Sze SK,
de Kleijn DPV, Lai RC, Tan EKY, Zhao H, Yeo KS,
Low TY, Lian Q, Lee CN, Mitchell W., El Oakley
RM, and Lim S-K(2007) Elucidating the secretion
proteome of human ESC-derived mesenchymal stem
cells. Molecular and Cellular Proteomics
61680-1689. 4. Que J, Lian Q, El Oakley RM,
Lim B, Lim S-K(2007) PI3 K/Akt/mTOR regulates
self-renewal and endothelial differentiation in
lineage restricted RoSH stem cell lines. J Mol
Signal. Sep 252(1)9 Epub ahead of print.