Isolation and characterization of mesenchymal stem cells derived from fetal bovine liver
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ORIGINAL PAPER
Isolation and characterization of mesenchymal stem cells derived from fetal bovine liver Taofeng Lu • Pengfei Hu • Xiaohua Su • Changli Li • Yuehui Ma • Weijun Guan
Received: 19 July 2013 / Accepted: 15 November 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract Bovine liver-derived mesenchymal stem cells (bLMSCs) were isolated from the liver tissue of 4–6 months old fetal calf, and then characterized by immunofluorescence and RT-PCR. We found that primary bLMSCs could be subcultured to 44 passages, the total culture time in vitro was 192 days. The results of surface antigen detection showed that bBMSCs expressed CD29, CD44, CD73, CD90, CD106 and CD166 but not expressed endothelial cells and hematopoietic cells specific marker CD34, CD45 and BLADR. The results of growth kinetics, colony-forming cell assay and cell cycle analysis indicated that the fetal bovine LMSCs had good proliferation ability in vitro. The cells from passages 7 were successfully induced to differentiate into osteoblasts, adipocytes and chondrocytes. The results indicate the potential for multi-lineage differentiation of bLMSCs that may represent an ideal candidate for cellular transplantation therapy. Keywords Bovine LMSCs Proliferation Multi-lineage differentiation
Taofeng Lu and Pengfei Hu have contributed equally to this work. T. Lu P. Hu X. Su C. Li Y. Ma W. Guan (&) Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China e-mail: [email protected]
Introduction Mesenchymal stem cells (MSCs) has the characteristics of self-renewal, immune regulation and pluripotent, they are an important member of the stem cell family, and they are extensively and deeply studied in recent years. MSCs were first found in the bone marrow, they could be isolated from fat (Campagnoli et al. 2001), liver (Campagnoli et al. 2001), peripheral blood (Huss et al. 2000), amniotic membrane (Moon et al. 2008), amniotic fluid (De Coppi et al. 2007); lung (Gong et al. 2012), synovium (De Bari et al. 2001), muscle (Yoshimura et al. 2007) and other fetal tissues. MSCs is mesoderm-derived pluripotent stem cell, it has a fiber-like shape, long-term survival and limited passage frequency, it also has the ability of selfrenewal and multi-lineage differentiation potential, it could be induced to differentiate into adipocytes (Fink et al. 2004), osteoblasts (Jaiswal et al. 1997), chondrocytes (Mackay et al. 1998; Barry et al. 2001; Worster et al. 2001), muscle cells (Wakitani et al. 1995), nerve cells (Black and Woodbury 2001; Sanchez-Ramos et al. 2000), and liver cells (Lee et al. 2004) and pancreatic b cells (Chen et al. 2004) in vivo and in vitro. However, MSCs derived from bone marrow and cord blood have been fully studied, while study on liver derived mesenchymal stem cells is rare. To date, there is no report on fetal derived bovine liver stem cells. In the normal state of the liver tissue, the cells are in the static status, only a few of them are in the
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