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To reduce the cost of cytokines and growth factors in stem cell research, a simple method for the production of soluble and biological active human basic fibroblast growth factor (hbFGF) fusion protein in Escherichia coli was established. Under optimal conditions, approximately 60-80 mg of >95% pure hbFGF fusion proteins (Trx-6xHis-hbFGF and 6xHis-hbFGF) were obtained from 1 liter of culture broth. The purified hbFGF proteins, both with and without the fusion tags, were biologically active, which was confirmed by their ability to stimulate proliferation of NIH3T3 cells. The fusion proteins also have the ability to support several culture passages of undifferentiated human embryonic stem cells and induce pluripotent stem cells. This paper describes a low-cost and uncomplicated method for the production and purification of biologically active hbFGF fusion proteins., (© 2015 S. Karger AG, Basel.)

Human Wharton's jelly–derived mesenchymal stromal cells (hWJ–MSCs) of umbilical cord tissue are a richest and more attractive source for stem cell–based therapy. Despite of their ability to undergo self–renewal, they also differentiate into all mesenchyme and some non–mesenchyme tissues, including hepatocytes. This study, we differentiated hWJ–MSCs into hepatocyte–like cells by using sodium butyrate (NaB) pre–treated with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) supplementation. Hepatic differentiation of hWJ–MSCs was induced with 3–step differentiation protocol. The differentiated cells were examined for the expression of hepatic–specific markers and hepatocyte function by using immunocytochemistry, real–time reverse transcriptase–polymerase chain reaction (RT–PCR) and Periodic acid–Schiff (PAS) staining. Before hepatic differentiation of hWJ–MSCs, we identified embryonic definitive endodermal cells by using RT–PCR, which increase of CXCR4 mRNA level, after NaB pre–treated along with EGF and bFGF supplementation. Hepatic differentiation of hWJ–MSCs by using NaB pre–treated along with EGF and bFGF supplementation, the differentiated hWJ–MSCs performed high hepatic–specific markers at gene and protein levels, and increased signal of glycogen storage accumulation. Our result indicate that hWJ–MSCs are able to differentiate into hepatocyte–like cells by inducing of hepatic differentiation medium and NaB pre–treated with EGF and bFGF supplementation. Therefore, the differentiated hWJ–MSCs to hepatocyte–like cells by inducing of NaB pre–treated with EGF and bFGF condition could represent an alternative source of end–stage of liver diseases. [ABSTRACT FROM AUTHOR]