无血清培养骺板细胞分泌TGF-β1的实验研究

TGF-β1 secreted by epiphyseal plate chondrocytes in serum-free medium

目的观察骺板细胞在无血清培养液中的生长情况,并通过ELISA方法检测骺板细胞分泌TGF-β1。方法提取3周龄新西兰兔骺板组织,获得良好生物活性的骺板细胞。采用CCK-8生长曲线检测骺板细胞在无血清培养液中的生长增殖情况,并通过ELISA方法检测骺板细胞分泌TGF-β1。结果骺板细胞生长情况良好,分泌细胞外基质。AO/PI荧光染色显示存活的骺板细胞呈亮绿色,周围散在有少量红染的死细胞。骺板细胞在DMEM培养液以及无血清培养液培养时生长曲线形态基本相似,在1、2d时细胞增殖缓慢,之后增殖速度逐渐增加,6-7d逐渐进入平台期。第1天时骺板细胞开始释放TGF-β1,然后释放量逐渐增加,第6d时浓度达到167.2pg/ml。结论骺板细胞在无血清培养液中增殖情况良好并能分泌TGF-β1。

Objective To observes the growth of epiphyseal plate chondrocytes in serum-free medium and detect the TGF-β1 secreted from epiphyseal plate chondrocyes by ELISA.Methods Epiphyseal plate chondrocytes were isolated from the epiphyseal tissue of New Zealand rabbits at the age of three weeks.Growth of epiphseal plate chondrocytes in serum-free medium was observed following the CCK-8 growth curve.The TGF-β1 secreted by epiphseal plate chondrocytes was detected by ELISA.Results The epiphseal plate chondrocyes grew quite well and secreted the extracellular matrix.AO/PI fluorescence staining showed bright green surviving cells with a small number of epiphseal plate chondrocyes red-stained around.The morphology of growth curves of the epiphseal plate chondrocyes in DMEM and serum-free medium was similar.The proliferation of epiphseal plate chondrocytes was slow on the first 2 days,then increased gradually,and entered the platform period on the 6-7 days.The epiphseal plate chondrocytes began to release the TGF-β1 on the first day,then increased to release it gradually,and the concentration of TGF-β1 reached 167.2pg/ml on the sixth day.Conclusion The epiphseal plate chondrocytes can grow well in serum-free medium and secrete TGF-β.