The three cytokines, hLIF, hbFGF, and mSCF, are known to retain the pluripotency and proliferation potential of human and mouse stem cells [35, 36].
In our experiment, stage X avian blastoderm cells with stem cell morphology and epitope profiles could be maintained with stem-like characteristics for a long term in vitro with the support of the DF-1 feeder and basic culture medium supplemented with only three cytokines: human basic fibroblast growth factor (hbFGF), mouse stem cell factor (mSCF), and human leukemia inhibitory factor (hLIF).
(c) The combination of 20ng/[micro]l each of hLIF, hbFGF, and mSCF was the optimal concentration.
In addition, stable protein expression of hbFGF was observed in MSCs after transduction of an adeno-associated virus vector (23).
In the present study, increased MSC survival was observed in those overexpressing hbFGF, which is consistent with previous studies reporting that hbFGF increased survival rate of transplanted stem cells (16), as well as proliferation of stem cells (22).
Moreover, although previous studies have successfully used hbFGF in rats (39,40), use of rat bFGF might have resulted in greater effectiveness.
Therefore, patients with ischemic disease may benefit from therapeutic angiogenesis mediated by a combination of gene therapy (i.e., hbFGF overexpression) with allogenic MSC transplantation.
Overexpression of human basic fibroblast growth factor (hbFGF) by mesenchymal stem cells (MSCs) in a rat hind limb ischemia model.
A, Representative images of green fluorescent protein (GFP) (left panels, green) and CD31 (middle panels, red) was determined by immunofluorescence analysis of the gastrocnemius muscle from the MSC, human basic fibroblast growth factor (hbFGF)-MSC, and GFP-MSC groups.