髓核间质干细胞在TGF-β1诱导下向类髓核细胞分化的实验研究

发布时间：2018-05-01 01:36

  本文选题：髓核间质干细胞 + 脂肪间质干细胞　； 参考：《山西医科大学》2015年硕士论文

【摘要】：目的:比较髓核间质干细胞与脂肪间质干细胞体外向类髓核细胞诱导分化能力的差异。方法:分别取大鼠腹股沟处脂肪组织与尾段脊柱,采用机械酶消化法分离培养AD-MSC与NP-MSC。流式细胞仪检测两种干细胞CD105、CD90、CD29、CD45、CD44、CD34、CD24的表达。AD-MSC与NP-MSC各自分为诱导组、无因子诱导组和对照组。诱导组以TGF-beta1标准软骨诱导液培养;无因子诱导组以不含TGF-beta1的软骨诱导液培养;对照组以含10%胎牛血清的DMEM/F12培养液培养。培养14d后用RT-PCR检测各组细胞Ⅱ型胶原、蛋白多糖、SOX-9基因的表达。结果:两种干细胞CD105、CD90、CD29表达阳性;CD45、CD44、CD34、CD24表达阴性。向类髓核细胞诱导培养14d后两种细胞诱导组Collagen typeⅡ、Aggrecan、SOX-9基因表达水平较对照组均明显升高,差异有统计学意义(P0.05);NP-MSC诱导组三种基因的表达水平均明显高于AD-MSC诱导组,差异有统计学意义(P0.05)。结论:髓核间质干细胞与脂肪间质干细胞体外均具有向类髓核细胞分化的能力,而髓核间质干细胞可能更优于脂肪间质干细胞,更适合于作为组织工程髓核研究的种子细胞。
[Abstract]:Aim: to compare the differentiation ability of nucleus pulposus mesenchymal stem cells and adipose mesenchymal stem cells into nucleus pulposus cells in vitro. Methods: AD-MSC and NP-MSCs were isolated from rat inguinal adipose tissue and caudal spine by mechanical enzyme digestion. Flow cytometry was used to detect the expression of CD105, CD90, CD29, CD45, CD44, CD34 and CD24 in stem cells. AD-MSC and NP-MSC were divided into three groups: induction group, non-factor induced group and control group. The induction group was cultured with TGF-beta1 standard cartilage induction medium, the non-factor induced group with TGF-beta1 free cartilage culture medium, and the control group with DMEM/F12 culture medium containing 10% fetal bovine serum. After cultured for 14 days, the expression of type 鈪，

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