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l:l,以脐带间充质于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.%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 cellbased therapies of degenerative disc disease.
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