| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 25502 | Journal of Biotechnology | 2006 | 21 Pages |
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-β2. Microarray analysis of immortalized and non-immortalized MEF revealed differential gene expression of several TGF-β related genes. By supplementing TGF-β2 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-β2) feeder cells including the induction of osteopontin following addition of TGF-β2. Our results demonstrate that TGF-β2 and its related genes in MEF play a role in the support of pluripotent hESC expansion.
