In vitro culture of bone marrow-derived mesenchymal stem cells under serum-free condition
The aim of this work is to establish a chemically-defined serum free medium (CDSFM) supports in vitro proliferation and maintains the multilineage differentiation potential of bone marrow-derived mesenchymal stem cells (BM-MSCs). BM-MSCs were isolated from the femoral bone of one-month-old New Zealand rabbits with combined density gradient centrifugation (Ficoll, γ 1.073) and attachment culture method. And compared the proliferation capability, cell cycle, colony-forming efficiency and osteogenic, adipogenic differentiation capability of BM-MSCs cultured in the CDSFM and serum-containing medium (SCM). Based on α -MEM, the SCM was supplied with 10% fetal bovine serum, the CDSFM was supplied with bFGF (10 mg/L), EGF (10 mg/L), bovine serum albumin ( BSA, 5 g/L ) , reduced glutathione ( 1.5 mg/L ) , β -mercaptoethanol ( 0.1 mM ) , SITE ( 1%, v/v ),dexamethasone ( 10nM ) and hydrocortisone (10 mg/L). The culture dishes were coating with rat tail collagen type I (0.1 mg/cm2). Over 10 days culture, resulting in 50-fold increases in cell number in the CDSFM compared to 40-fold in the SCM. bFGF, EGF and hydrocortisone were the most important additives that significantly stimulated BM-MSCs proliferation. The percentage of cells at G0-G1 cell cycle was (80.31±0.58)% in the CDSFM compare to (75.24±4.05)% in the SCM without significant difference (P>0.05). However, the cloning efficiency of BM-MSCs in the CDSFM was lower than that in the SCM with significant difference (p<0.01). The expanded MSCs in the CDSFM preserved differentiation potentials into mesenchymal lineages in vitro, including adipocytes and osteoblasts. A chemically-defined serum free medium was designed to support in vitro proliferation and maintains the stem cells characters of BM-MSCs, which could be applied to cell-based therapy and biomedical research.
bone marrow-derived mesenchymal stem cells serum free medium proliferation multilineage differentiation tissue engineering
Wei Wu Yan Zhou Wen-Song Tan
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology,Shanghai 200237, PR China.
国际会议
The 12th Asian Pacific Confederation of Chemical Engineering Congress(第十二届亚太化工联盟大会暨化工展览会)
大连
英文
2008-08-04（万方平台首次上网日期，不代表论文的发表时间）