Growth factor expression pattern of homologous feeder layer for culturing buffalo embryonic stem cell-like cells
Ruchi Sharma A , Aman George A , Nitin M. Kamble A , Manmohan S. Chauhan A , Suresh Singla A , Radhey S. Manik A and Prabhat Palta A B
+ Author Affiliations
- Author Affiliations
A Embryo Biotechnology Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal-132001, Haryana, India.
B Corresponding author. Email: prabhatpalta@yahoo.com
Reproduction, Fertility and Development 24(8) 1098-1104 https://doi.org/10.1071/RD11298
Submitted: 1 December 2011 Accepted: 23 March 2012 Published: 24 April 2012
Abstract
The present study examined the expression profile of buffalo fetal fibroblasts (BFF) used as a feeder layer for embryonic stem (ES) cell-like cells. The expression of important growth factors was detected in cells at different passages. Mitomycin-C inactivation increased relative expression levels of ACTIVIN-A, TGF-β1, BMP-4 and GREMLIN but not of fibroblast growth factor-2 (FGF-2). The expression level of ACTIVIN-A, transforming growth factor-β1 (TGF-β1), bone morphogenetic protein-4 (BMP-4) and FGF-2 was similar in buffalo fetal fibroblast (BFF) cultured in stem cell medium (SCM), SCM + 1000 IU mL–1 leukemia inhibitory factor (LIF), SCM + 5 ng mL–1 FGF-2 or SCM + LIF + FGF-2 for 24 h whereas GREMLIN expression was higher in FGF-2-supplemented groups. In spent medium, the concentration of ACTIVIN-A was higher in FGF-2-supplemented groups whereas that of TGF-β1 was similar in SCM and LIF + FGF-2, which was higher than when either LIF or FGF-2 was used alone. Following culture of ES cell-like cells on a feeder layer for 24 h, the TGF-β1 concentration was higher with LIF+FGF-2 than with LIF or FGF-2 alone which, in turn, was higher than that in SCM. In the LIF + FGF-2 group, the concentration of TGF-β1 was lower and that of ACTIVIN-A was higher in spent medium at 24 h than at 48 h of culture. These results suggest that BFF produce signalling molecules that may help in self-renewal of buffalo ES cell-like cells.
Additional keywords: ACTIVIN-A, fetal fibroblast, mitomycin-C, pluripotency, TGF-β1.
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