Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro |
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| Authors: | CAI Yun-feng MIN Jun FANG Tian-ling CHU Zhong-hua DENG Xiao-geng HE Jing-song CHEN Ji-sheng |
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| Institution: | 1. Department of Hepatic-Biliary Surgery, The 2nd Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, China;
2. Stem Cell Research Center, The 2nd Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, China;
3. Research Center of Medicine, The 2nd Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, China |
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| Abstract: | AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy. |
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| Keywords: | Bone marrow Stem cells Cell culture Cell separation |
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