Growth and Propagation of Differentiated Cells Derived From Embryonic Stem (ES) Like Cells of <i>Cirrhinus mrigala </i>(Ham.)

Abstract

In this study, we describe the growth and propagation of differentiated cells viz. keratinocyte-like cells (ectodermal origin), muscle-like cells (mesodermal origin) and hepatocyte-like cells (endodermal origin) generated from embryoid bodies (EBs) in their respective formulated medium. These EBs were developed by culturing mrigal ES-like cells in suspension. When these EBs were attached to a permissive surface and upon treatment with appropriate inducer like retinoic acid (RA) they continued to differentiate into different cell lineages. Here, we formulated three specific growth and propagation media for these derived differentiated cells. The mediums were supplemented with key factors required for their growth and propagation. Keratinocyte growth and propagation medium (KGPM) was supplemented with 5 ng/ml keratinocytes growth factor (KGF), 5 ng/ml epidermal growth factor (EGF), 5 μg/ml insulin and optimum calcium concentration (0.05 mM) for keratinocyte-like cells. Similarly, 10 ng/mL EGF, 0.5 mg/mL BSA and 100 ng/mL insulin-like growth factor -1 (IGF-1) were supplemented in muscle cells growth and propagation medium (MGPM) for muscle-like cells and 10 ng/ml EGF along with 2% dimethyl sulfoxide (DMSO) was supplemented in hepatocyte growth and propagation medium (HGPM) for hepatocyte-like cells.