Human collagen-I has been widely used as coating material for various cell culture and specific lineage differentiation application. To provide a defined coating matrix for potential therapeutic development, various functional domain of collagen-I has been identified. One of domain (124 - 402aa, CBF8) was demonstrated to be hepatocytes specific attachment matrix through a3b1 integrin mediated adhesion. Human vitronectin (VTN) is a liver derived major plasma glycoprotein that exhibits multiple activities and functions as a cell adhesion molecule and regulator of coagulation. It contains the amino acid structural motif, Arg-Gly-Asp (RGD), which is involved in cell attachment. Recombinant human VTN protein has been successfully used as coating matrix protein for long-term culture of human ES cells with excellent binding affinity for polystyrene culture dishes. Purified human VTN- CBF8 fusion protein may provides an ideal coating matrix protein for in vitro expansion of human Hepatocytes.

Human collagen-I CBF8 domain cDNA fragment (26 - 542 aa) was constructed with N-terminal fusion of human VTN (61-398aa) and expressed in E.coli as inclusion bodies. The final product was refolded using our unique “temperature shift inclusion body refolding” technology and chromatographically purified as soluble protein. Coating this recombinant protein at 5-10 ug / well (6 well plate) in Hepatocytes specific medium can be used for human liver cell differentiation study in vitro.

1. May be used for in vitro human hepatocytes differentiation ascoating matrix protein.

2. May be used as culture matrix protein for long term liver cells cultivation in vitro.