自体骨髓间充质干细胞和同种异体软骨细胞共培养优化软骨组织工程种子的细胞源

摘要

BACKGROUND: There have been no cells that can meet the requirement of tissue engineering for seed cellsOBJECTIVE: To investigate the feasibility of co-culturing autogenic bone marrow mesenchymal stem cells and allogenicchondrocytes.METHODS: Rabbit chondorcytes were harvested by enzymatic digestion Bone marrow mesenchymal stem cells were harvestedby density gradient centrifugation and adherence methods Passage 2 bone marrow mesenchymal stem cells and chondrocytesat a concentration of 3Χ108/L were randomly dicided imto three groups.Co-culture group:bone marrow mesenchymal stem cellswere co-cultured with chondrocytes at a ratio of 2: 1 Experimental group Chondrocytes of the same passage and concentration(concentration the same as the final concentration of co-culture group):control group:1Χ108/Lchondrocytes(concentration thesame as the final concentration of co-culture group).RESULTS AND CONCLUSION: The average population doubling time was 3 days in the co-culture group, 7 days in the experimental group, and 8 days in the control group Cell proliferation was significantly faster in the co-culture group than in the experimental and control groups (P < 0 05) The glycosaminoglycan level was significantly higher in the co-culture group than in the experimental and control groups (P < 0 05) There was no obvious rejection in the co-cultures of autogenic bone marrowmesenchymal promote the proliferation of chondrocytes.This of extracellular matrix.shorten culture time ofcultures can promote the proliferation of chondrocytes and the synthesis of extracellular matrix, shorten culture time of chondrocytes, and reduce passages, at the same time, chondrocytes can promote oriented transformation of bone marrow mesenchymal stem cells towards chondrocytes, which save a large number of chondrocytes%背景:至今尚无一种细胞能完全满足组织工程对种子细胞的要求.目的:探讨自体骨髓间充质干细胞和同种异体软骨细胞共培养的可行性.方法:酶消化分离法获得兔软骨细胞,使用密度梯度离心和贴壁筛选的方法获得骨髓间充质干细胞.取细胞浓度为3×108 L-1的第2代骨髓间充质干细胞和软骨细胞,随机分为3组,共培养组:将骨髓间充质干细胞和软骨细胞按2∶1比例混匀;实验组:取同代同浓度的软骨细胞(浓度与共培养细胞的终浓度相同);对照组:取低浓度软骨细胞1×108 L-1(与共培养组中软骨细胞终浓度相同).结果与结论:共培养组、实验组及对照组细胞平均群体倍增时间分别为3,7,8 d;共培养组共培养细胞增殖比其他2组明显增快(P < 0.05);糖胺多糖水平明显多于其他2组(P < 0.05);自体骨髓间充质干细胞与同种异体软骨细胞共培养未见明显排异反应,提示自体骨髓间充质干细胞与同种异体软骨细胞为种子细胞共培养,骨髓间充质干细胞能促进软骨细胞的增殖和细胞外基质合成,缩短软骨细胞培养时间和减少传代次数,同时软骨细胞可促进骨髓间充质干细胞向软骨细胞的定向转化,节省大量软骨细胞.