title: Growth factor expression pattern of homologous feeder layer for culturing buffalo embryonic stem cell-like cells.
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+1000IU mL(-1) leukemia inhibitory factor (LIF), SCM+5 ngmL(-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.
authors.affiliation: Embryo Biotechnology Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal-132001, Haryana, India.
authors.forename: Ruchi
authors.initials: R
authors.lastname: Sharma
authors.forename: Aman
authors.initials: A
authors.lastname: George
authors.forename: Nitin M
authors.initials: NM
authors.lastname: Kamble
authors.forename: Manmohan S
authors.initials: MS
authors.lastname: Chauhan
authors.forename: Suresh
authors.initials: S
authors.lastname: Singla
authors.forename: Radhey S
authors.initials: RS
authors.lastname: Manik
authors.forename: Prabhat
authors.initials: P
authors.lastname: Palta
mesh_terms: D028341:Activins / Q000032:analysis / Q000235:genetics; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000032:analysis / Q000235:genetics; D019485:Bone Morphogenetic Proteins / Q000032:analysis / Q000235:genetics; D002020:Buffaloes / Q000196:embryology*; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary*; D017077:Culture Media, Conditioned / Q000737:chemistry*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling / Q000662:veterinary*; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000235:genetics*; D011247:Pregnancy; D053773:Transforming Growth Factor beta1 / Q000032:analysis
chemical_list: D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D017077:Culture Media, Conditioned; D036341:Intercellular Signaling Peptides and Proteins; D053773:Transforming Growth Factor beta1; C422405:activin A; D028341:Activins