Generation of buffalo embryonic stem cell clones on matrigel and fibronectin synthetic matrices and their characterization

Abstract

The present experiment was carried out to generate embryonic stem cell clones on matrigel and fibronectin synthetic matrices and their characterization in buffalo. In vitro derived buffalo embryos were made zona free and were cultured on matrigel and fibronectin synthetic matrices with 4 culture media, viz. control media: media 1: CM + SCF; media 2: M-I + bFGF; and media 3: M-II + IGF -1 in CO₂ incubator. A total of 16 numbers of zona free embryos were cultured
in each culture media on both the synthetic matrices and the efficiency of clumped blastomere attachment, stem cell
clones formation and their propagation was observed. The primary stem cell clone formation in culture media 1 and 3
both were found to be significantly higher than media–2 and control. The embryos attachment percentages on both the
matrices were also better in M-1and M-3 as compare to media-2 and control.The results showed that embryonic stem
cell proliferation was better in both SCF supplemented media and combination with IGF for lineage maintenance. It
was also observed that the stem cell colony formation was better on matrigel than fibronectin matrices which may be
due to the fact that the pluripotent gene expression was better in matrigel based culture than the fibronectin synthetics. The clones formed in all the medium expressed pluripotent markers vide Oct4, Nanog, Klf4 and Foxd3 which implies that the stem cell clones were stem cell like cells. It can be concluded that buffalo ES cell can be established on
synthetic matrices.