非接触式共培养体系对脐带间充质干细胞向类髓核细胞的诱导分化效应

目的探讨人脐带间充质干细胞向类髓核细胞的分化潜能,为椎间盘退变性疾病的生物学治疗探索新的细胞来源。方法取人正常足月产婴儿脐带及成人椎间盘,分离消化华通胶组织和髓核组织,收集培养脐带间充质干细胞及成人髓核细胞。利用带有插入层的Transwell 6孔培养板进行细胞非接触式共培养,细胞比例为1∶1,以脐带间充质干细胞单独培养作为对照组。共培养1周后,提取脐带间充质干细胞总RNA,利用Real-Time PCR检测其Ⅱ型胶原、蛋白多糖及SOX9基因的相对表达改变。结果成功分离提取脐带间充质干细胞;Real-Time PCR检测结果显示实验组脐带间充质干细胞Ⅱ型胶原、蛋白多糖和SOX9基因相对表达较对照组显著增加,差异有统计学意义（P〈0.01）。结论人脐带间充质干细胞能够被髓核细胞诱导分化为类髓核细胞,有可能为椎间盘退变性疾病的生物学治疗提供一种新的细胞来源。

Effect of indirect co-culture system on differentiation of human umbilical cord-derived mesenchymal stem cells into nucleus pulposus-like cells

Objective To evaluate the differentiation potential of human umbilical cord-derived mesenchymal stem cells（UCMSCs） into nucleus pulposus-like cells（NPCs）,so as to search for a new cell source for biotherapy of degenerative disc disease.Methods Umbilical cord was obtained after normal birth and the mesenchyamal stem cells were isolated from the umbilical cord Wharton＇s jelly.Transwell six-well plates were used for coculture without contact,with the cells seeded at a ratio of 1∶1.UCMSCs and NPCs were co-cultured for 7 d.UCMSCs cultured alone served as controls.The total RNA was extracted from cells using Trizol reagent and Real-time PCR was used to examine the expression of type II collagen,aggrecan,and SOX9.Results UCMSCs were successfully isolated from human umbilical cord.Real-time PCR results showed that SOX9,type II collagen and aggrecan mRNA were significantly increased in the coculture group compared with that in the control group（P〈0.01）.Conclusion It has been found that coculture of NPCs with UCMSCs can induce UCMSCs differentiating into an NPCs phenotype,indicating that UCMSCs might be a promising seed cells for tissue engineering therapies or cell-based therapies of degenerative disc disease.