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World J Gastroenterol. Apr 7, 2009; 15(13): 1630-1635
Published online Apr 7, 2009. doi: 10.3748/wjg.15.1630
Passage of bone-marrow-derived liver stem cells in a proliferating culture system
Yun-Feng Cai, Ji-Sheng Chen, Shu-Ying Su, Zuo-Jun Zhen, Huan-Wei Chen
Yun-Feng Cai, Shu-Ying Su, Zuo-Jun Zhen, Huan-Wei Chen, Department of Hepatobiliary Surgery, the Affiliated Foshan Hospital of Sun Yat-sen University, Foshan 528000, Guangdong Province, China
Ji-Sheng Chen, Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, Guangdong Province, China
Author contributions: Cai YF and Chen JS contributed equally to this work; Cai YF, Chen JS, Su SY, Zhen ZJ and Chen HW designed the research; Cai YF, Su SY, Zhen ZJ and Chen HW performed the research; Chen HW contributed new reagents/analytic tools; Cai YF, Chen JS and Zhen ZJ analyzed the data; Cai YF, Chen JS, Chen HW wrote the paper.
Correspondence to: Yun-Feng Cai, MD, Department of Hepatobiliary Surgery, the affiliated Foshan Hospital of Sun Yat-sen University, Foshan 528000, Guangdong Province, China. yfcai70@yahoo.com.cn
Telephone: +86-757-83161119
Fax: +86-757-83812566
Received: December 24, 2008
Revised: March 9, 2009
Accepted: March 16, 2009
Published online: April 7, 2009
Abstract

AIM: To explore the feasibility of passage of bone-marrow-derived liver stem cells (BDLSCs) in culture systems that contain cholestatic serum.

METHODS: Whole bone marrow cells of rats were purified with conditioning selection media that contained 50 mL/L cholestatic serum. The selected BDLSCs were grown in a proliferating culture system and a differentiating culture system. The culture systems contained factors that stimulated the proliferation and differentiation of BDLSCs. Each passage of the proliferated stem cells was subjected to flow cytometry to detect stem cell markers. The morphology and phenotypic markers of BDLSCs were characterized using immunohistochemistry, reverse transcription polymerase chain reaction (RT-PCR) and electron microscopy. The metabolic functions of differentiated cells were also determined by glycogen staining and urea assay.

RESULTS: The conditioning selection medium isolated BDLSCs directly from cultured bone marrow cells. The selected BDLSCs could be proliferated for six passages and maintained stable markers in our proliferating system. When the culture system was changed to a differentiating system, hepatocyte-like colony-forming units (H-CFUs) were formed. H-CFUs expressed markers of embryonic hepatocytes (alpha-fetoprotein, albumin and cytokeratin 8/18), biliary cells (cytokeratin 19), hepatocyte functional proteins (transthyretin and cytochrome P450-2b1), and hepatocyte nuclear factors 1α and -3β). They also had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes.

CONCLUSION: BDLSCs can be selected directly from bone marrow cells, and pure BDLSCs can be proliferated for six passages. The differentiated cells have hepatocyte-like phenotypes and functions. BDLSCs represent a new method to provide a readily available alternate source of cells for clinical hepatocyte therapy.

Keywords: Liver stem cells; Bone marrow; Cell separation; Cell proliferation; Cell differentiation