体外软骨细胞与软骨单位共培养构建软骨组织工程种子细胞的最佳比值

发布时间：2018-03-24 05:34

本文选题：软骨细胞　切入点：软骨单位　出处：《山西医科大学》2013年硕士论文

【摘要】：目的：利用兔关节软骨细胞与软骨单位建立混合共培养体系，明确体外软骨细胞与软骨单位共培养构建软骨组织工程种子细胞的最佳比值。方法：将兔膝关节软骨分别用酶消化为软骨细胞与软骨单位后，按一定比例混合配制，分为五组，A组:单纯软骨细胞组；B组:比值为2:1的软骨细胞/软骨单位组；C组:比值为1:1的软骨细胞/软骨单位组；D组:比值为1:2的软骨细胞/软骨单位组；E:单纯软骨单位组。以2×105个细胞/孔接种于放有盖玻片的六孔板中，使细胞单层贴壁培养爬片，待细胞生长6日爬满后，进行指标检测： HE、番红O染色观察细胞组织形态学的变化，RT-PCR检测Agg、II型胶原和I型胶原的基因表达水平。Elisa检测上清液GAG、II型胶原的含量。结果：共培养组细胞形态正常。通过RT-PCR检测Agg、II型胶原和I型胶原的基因表达，Elisa检测上清液GAG、II型胶原的含量表明：采用共培养方法的软骨细胞与软骨单位的混合组中细胞代谢的表达要明显高于单纯软骨细胞组，尤其在软骨细胞/软骨单位共培养组（1:1），糖胺多糖与II型胶原在基因的表达和代谢情况均为最高。结论：在软骨组织工程移植种子细胞时，，软骨细胞与软骨单位为1∶1的比例为最佳选择，适用于软骨组织工程。
[Abstract]:Aim: to establish a mixed co-culture system of rabbit articular chondrocytes and chondrocytes to determine the optimal ratio of chondrocytes co-cultured with chondrocytes to chondrocyte seed cells for cartilage tissue engineering. Methods: the articular cartilage of rabbit knee joint was digested into chondrocytes and chondrocytes by enzyme, then mixed into chondrocytes and chondrocytes in a certain proportion. Divided into five groups: chondrocyte group B: chondrocyte / chondrocyte unit group C group with 2:1; chondrocyte / chondrocyte unit group D group with 1:1; chondrocyte / chondrocyte / chondrounit group with 1:2 ratio: single chondrocyte / chondrocyte unit group. Pure chondrocyte unit group. Inoculated with 2 脳 105 cells / hole in a six-hole plate with a cover glass, Make the cell monolayer adhere to the wall of the climbing piece, after 6 days of cell growth full, The changes of histomorphology were observed by HEand Phangnon O staining. The gene expression level of type II and type I collagen was detected by RT-PCR. Elisa was used to detect the content of type II collagen in supernatant. Results: the morphology of the co-cultured cells was normal. The gene expression of type II collagen and type I collagen was detected by RT-PCR. The content of type II collagen in supernatant was determined by Elisa. The results showed that the chondrocytes and chondrocytes were mixed with chondrocytes by co-culture method. The expression of cell metabolism in the chondrocyte group was significantly higher than that in the chondrocyte group. Especially in chondrocyte / chondrocyte coculture group (1: 1), glycosaminoglycan and type II collagen had the highest gene expression and metabolism. Conclusion: the ratio of chondrocytes to chondrocytes is 1:1, which is suitable for cartilage tissue engineering.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R329

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