Selective proliferation of rat hepatocyte progenitor cells in serum-free culture

Abstract

This protocol details a method of obtaining selectively proliferated hepatocyte progenitor cells using hyaluronic acid (HA)-coated dishes and serum-free medium. A small hepatocyte (SH) is a hepatocyte progenitor cell of adult livers and has many hepatic functions. When the rat SH begins to proliferate, CD44 is specifically expressed. To define the purification of SH, CD44 and cytokeratin 8 are used as marker proteins. The growth of SHs is faster on HA-coated dishes than on other extracellular matrix-coated ones. The use of both DMEM/F12 medium and HA-coated dishes allows the selective proliferation of SHs in culture. The purification of SHs is approximately 85% at day 10.

The Role of the Sodium-Taurocholate Cotransporting Polypeptide (NTCP) and of the Bile Salt Export Pump (BSEP) in Physiology and Pathophysiology of Bile Formation

“…This relatively late appearance of Ntcp during ontogenesis is reproduced in primary cultures of small hepatocytes, which expand and differentiate into mature hepatocytes [243,280]. Small hepatocytes are hepatocyte progenitor cells found in adult liver [44]. In primary cultured rat hepatocytes, Ntcp is rapidly down-regulated at the mRNA and at the protein level [195,261].…”

Thy1-Positive Cells Have Bipotential Ability to Differentiate into Hepatocytes and Biliary Epithelial Cells in Galactosamine-Induced Rat Liver Regeneration

“…25 After the perfusion, the cell suspension was centrifuged at 50 ϫ g for 1 minute. The supernatant and the precipitate were used for sorting Thy1 ϩ and CD44 ϩ cells and preparing MHs, respectively.…”

Culture of newborn monkey liver epithelial progenitor cells in chemical defined serum-free medium

“…Moreover, to ensure more reproducible culture techniques and enhanced safety, it is also indispensable to develop such culture system if cells will be applied to human cell therapy. However, only few studies employed chemical defined systems for long-time extensive expansion of HPCs (Tsuchiya et al 2005;Chen et al 2007;Turner et al 2007;Sasaki et al 2008;Arends et al 2009). …”