TGF-β2 allows pluripotent human embryonic stem cell proliferation on E6/E7 immortalized mouse embryonic fibroblasts

被引：18
作者：
Chen, S [1 ]
Choo, A [1 ]
Chin, A [1 ]
Oh, SKW [1 ]
机构：
[1] Bioproc Technol Inst, Stem Cell Grp, Singapore 138668, Singapore
来源：
JOURNAL OF BIOTECHNOLOGY | 2006年 / 122卷 / 03期
关键词：
TGF-beta; 2; E6/E7; osteopontin;
D O I：
10.1016/j.jbiotec.2005.11.022
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
In this study we report observations that mouse embryonic fibroblasts (MEF) capable of supporting expansion of pluripotent, human embryonic stem cells (hESC) fail to support after immortalization using E6/E7 oncogenes in serum conditions; however this can be reversed following addition of exogenous TGF-beta 2. Microarray analysis of immortalized and non-immortalized MEF revealed differential gene expression of several TGF-beta related genes. By supplementing TGF-beta 2 into E6/E7 immortalized MEF cultures, this enabled proliferation of undifferentiated, pluripotent hESC as demonstrated by marker expression (Oct-4, SSEA-4, alkaline phosphatase) and teratoma formation representing three germ layers following hESC injection into immuno-deficient mice. Subsequent investigation using quantitative real-time PCR highlighted differential gene expression of several extracellular matrix related transcripts in primary and immortal (+/- TGF-beta 2) feeder cells including the induction of osteopontin following addition of TGF-beta 2. Our results demonstrate that TGF-beta 2 and its related genes in MEF play a role in the support of pluripotent hESC expansion. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码：341 / 361
页数：21
相关论文
共 34 条
[1]   Feeder layer- and serum-free culture of human embryonic stem cells [J].
Amit, M ;
Shariki, C ;
Margulets, V ;
Itskovitz-Eldor, J .
BIOLOGY OF REPRODUCTION, 2004, 70 (03) :837-845
[2]   Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers [J].
Beattie, GM ;
Lopez, AD ;
Bucay, N ;
Hinton, A ;
Firpo, MT ;
King, CC ;
Hayek, A .
STEM CELLS, 2005, 23 (04) :489-495
[3]   THE E6-E7 REGION OF HUMAN PAPILLOMAVIRUS TYPE-18 IS SUFFICIENT FOR TRANSFORMATION OF NIH-3T3 AND RAT-1 CELLS [J].
BEDELL, MA ;
JONES, KH ;
LAIMINS, LA .
JOURNAL OF VIROLOGY, 1987, 61 (11) :3635-3640
[4]   Assessment of stem cell markers during long-term culture of mouse embryonic stem cells [J].
Berrill, A ;
Tan, HL ;
Wuang, SC ;
Fong, WJ ;
Choo, ABH ;
Oh, SKW .
CYTOTECHNOLOGY, 2004, 44 (1-2) :77-91
[5]   New POU dimer configuration mediates antagonistic control of an osteopontin preimplantation enhancer by Oct-4 and Sox-2 [J].
Botquin, V ;
Hess, H ;
Fuhrmann, G ;
Anastassiadis, C ;
Gross, MK ;
Vriend, G ;
Schöler, HR .
GENES & DEVELOPMENT, 1998, 12 (13) :2073-2090
[6]   REGULATION OF GROWTH AND GENE-EXPRESSION IN HUMAN PAPILLOMAVIRUS-TRANSFORMED KERATINOCYTES BY TRANSFORMING GROWTH-FACTOR-BETA - IMPLICATIONS FOR THE CONTROL OF PAPILLOMAVIRUS INFECTION [J].
BRAUN, L ;
DURST, M ;
MIKUMO, R ;
CROWLEY, A ;
ROBINSON, M .
MOLECULAR CARCINOGENESIS, 1992, 6 (02) :100-111
[7]   Expansion of pluripotent human embryonic stem cells on human feeders [J].
Choo, ABH ;
Padmanabhan, J ;
Chin, ACP ;
Oh, SKW .
BIOTECHNOLOGY AND BIOENGINEERING, 2004, 88 (03) :321-331
[8]   Transforming growth factor-beta (TGF-β) [J].
Clark, DA ;
Coker, R .
INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 1998, 30 (03) :293-298
[9]   Osteopontin as a means to cope with environmental insults: regulation of inflammation, tissue remodeling, and cell survival [J].
Denhardt, DT ;
Noda, M ;
O'Regan, AW ;
Pavlin, D ;
Berman, JS .
JOURNAL OF CLINICAL INVESTIGATION, 2001, 107 (09) :1055-1061
[10]   Direct binding of Smad3 and Smad4 to critical TGFβ-inducible elements in the promoter of human plasminogen activator inhibitor-type 1 gene [J].
Dennler, S ;
Itoh, S ;
Vivien, D ;
ten Dijke, P ;
Huet, S ;
Gauthier, JM .
EMBO JOURNAL, 1998, 17 (11) :3091-3100
1234