Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (40): 7425-7429.doi: 10.3969/j.issn.1673-8225.2010.40.004

Previous Articles     Next Articles

Culture and osteogenic differentiation of different-aged children bone marrow mesenchymal stem cells in autologous serum in vitro

Sun Ke1, Wang Guo-bing2, Zu Ying2, Tang Sheng-ping1, Yu Wei3, Chen Xiao-wen2　   

  1. 1 Department of Orthopaedics, 2 Institute of Pediatric Surgery, 3 Department of Ultrasound, Shenzhen Children Hospital, Shenzhen  518026, Guangdong Province, China
  • Online:2010-10-01 Published:2010-10-01
  • About author:Sun Ke★, Master, Associate chief physician, Department of Orthopaedics, Shenzhen Children Hospital, Shenzhen 518026, Guangdong Province, China soknow@126.com
  • Supported by:

     the Science and Technology Program of Shenzhen City, No. 200803058*

PDF

308

Abstract:

BACKGROUND: Most present studies emphasized on the induced osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) isolated from adults in vitro, but little was known about the osteogenic differentiation of hBMSCs isolated from children with different ages, or its biological characteristics after induced with autologous serum and osteogenic differentiation in vitro.
OBJECTIVE: To study culture conditions and methods of different-aged children BMSCs with autologous serum of in vitro culture, and explore its basic biological characteristics after osteogenic differentiation.
METHODS: Bone suspension of children, which were divided into three groups according to their age, was collected under aseptic conditions. hBMSCs were isolated and cultivated with media containing autologous serum. The cell growth curve was determined, and expression of cell surface antigens was analyzed by flow cytometry. After differentiation induced by β-sodium glycerophosphate, antiscorbic acid and dexamethasone, formation of alkaline phosphatase, type I collagen and calcium nodules of sub-cultivation cells were accessed by immunohistochemical method. The differentiation efficiency was evaluated. Growth conditions and morphological characteristics of primary cells, passage cells and differentiated cells were monitored using an inverted phase contrast microscope day by day.
RESULTS AND CONCLUSION: Among surface antigens cultivated hBMSCs from the three groups, CD29 and CD44 were detected to be positive, while CD34, CD45, CD105, CD106 and HLA-DR were found to be all negative. No significant difference of expression intensities were observed among three groups. The growth rate and differentiation efficiency of children BMSCs decreased with aging. Comparative studies on growth characteristics, surface antigen expression, differentiation potential of children hBMSCs suggested that autologous serum is a safe and effective culture method applicable to clinic with a high differentiation efficiency at the low child ages.

CLC Number: