微载体旋转立体培养成人退变髓核细胞增殖及生长动力学研究Growth and proliferation in adult degenerative nucleus pulposus cells with microcarrier culture in vitro
宁斌;刘海飞;龚维明;田庆芬;王德春;胡有谷;
摘要(Abstract):
[目的]通过单层培养和微载体旋转立体培养成人退变髓核细胞对比,研究两种培养方式对其生长活性及增殖能力的影响。[方法]本实验通过Trypan blue法计数细胞并绘制生长曲线,应用3H-TdR放射免疫法检测退变髓核细胞增殖活性,流式细胞仪检测退变髓核细胞周期,比较两种培养方式对细胞增殖的影响。[结果]微载体旋转立体旋转培养较单层培养细胞从贴壁到对数生长期开始时间(16~20 d)较长,但是严格意义的稳定生长期(3~5 d)很短;对数生长期长,一般14~17 d,期间细胞增殖活跃。两种培养方式的细胞处于稳定生长期时,细胞增殖能力没有明显区别,而处于对数生长期时,微载体培养的细胞的增殖能力明显高于单层培养组的细胞(P<0.05)。细胞周期测定显示两种细胞停滞在G0/G1期的占71%左右,培养方式对其影响不明显。而微载体培养的细胞的S期明显较单层培养的细胞为长(P<0.05)。[结论]成人退变髓核应用微载体培养后增殖能力及生长活性提高,可解决组织工程学椎间盘种子细胞难以收集的难题,为将来开展椎间盘组织工程学的研究奠定良好的基础。
关键词(KeyWords): 微载体;髓核细胞;细胞周期;生长曲线;细胞增殖
基金项目(Foundation): 国家自然基金面上项目资助(编号:30672132)
作者(Author): 宁斌;刘海飞;龚维明;田庆芬;王德春;胡有谷;
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