滑膜间充质干细胞与软骨细胞接触式共培养构建软骨的初步研究

doi:10.13701/j.cnki.kqyxyj.2016.06.002

口腔医学研究 ›› 2016, Vol. 32 ›› Issue (6): 554-558.DOI: 10.13701/j.cnki.kqyxyj.2016.06.002

滑膜间充质干细胞与软骨细胞接触式共培养构建软骨的初步研究

邵博¹ ,胡露露¹ ,龚忠诚^1* ,杨萌¹ ,宁晓婷¹ ,王玥¹ ,胡鑫¹ ,刘慧¹
克热木·阿巴斯¹ ,凌彬¹ ,尹小朋¹ ,高芸竹² ,林兆全¹

1. 新疆医科大学第一附属医院颌面肿瘤外科,新疆医科大学口腔医学院,新疆维吾尔自治区口腔医学研究所新疆乌鲁木齐 830054;
2. 新疆医科大学第一附属医院中心手术室新疆乌鲁木齐 830054

收稿日期:2015-11-19 出版日期:2016-06-26 发布日期:2016-06-22
通讯作者: 龚忠诚,电话:13319822466
作者简介:邵博(1986~ ),男,新疆人,住院医师,硕士,主要从事软骨组织工程研究工作。
基金资助:
国家自然科学基金项目(编号:31260229)
新疆维吾尔自治区优秀青年科技创新人才培养项目(编号:2014721046)

Direct Co-culture of Synovial Stem Cells and Chondrocytes Induces Enhanced Cartilage Tissue Formation.

SHAO Bo¹, HU Lu-lu¹, GONG Zhong-cheng^1*, YANG Meng¹, NING Xiao-ting¹, WANG Yue¹, HU Xin¹, LIU Hui¹, KEREMU·ABAShi¹, LIN Bin¹, YING Xiao-peng¹, GAO Yun-zu², LIN Zao-quan¹.

1. Department of Oral and Maxillofacial Surgery, School of Stomatology, Xinjiang Medical University. Urumqi 830054, China;
2. Centre Operation Room, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054

Received:2015-11-19 Online:2016-06-26 Published:2016-06-22

摘要/Abstract

摘要： 目的:探讨大鼠软骨细胞与滑膜间充质干细胞以1∶1比例接触式共培养,评价软骨细胞诱导干细胞在体外成软骨方向分化。方法:相同培养基体外培养大鼠滑膜间充质干细胞与软骨细胞,扩增至第3代滑膜间充质干细胞与第1代软骨细胞按1∶1比例混匀,以1×10⁶/mL为终浓度的微团体外培养作为实验组,以相同终浓度细胞数的软骨细胞和滑膜间充质干细胞作为对照组,每组各接种6组。各组标本于体外培养21d后,通过形态学观察,组织学染色,RT-PCR检测产物等方法对其新生软骨进行评价。结果:实验组及对照组体外培养21d后,形成微团似软骨样组织,质地较韧,乳白色。免疫组化法进行Ⅱ型胶原鉴定,基质能被Ⅱ型胶原染色。RT-PCR结果显示诱导后的微团表达软骨特异性基因Ⅱ型胶原和蛋白聚糖。结论:滑膜间充质干细胞与软骨细胞通过接触式共培养形成较成熟的软骨。

关键词: 滑膜间充质干细胞, 软骨细胞, 共培养, 组织工程

Abstract: Objective: To discuss direct co-culture of rat chondrocytes and synovial-derived mesenchymal stem cells at 1:1 and evaluate chondrocytes induced differentiation of stem cells in vitro. Methods: Rat synovial-derived mesenchymal stem cells and chondrocytes were cultivated in vitro in the same medium. The 3rd generation of synovial-derived mesenchymal stem cells and the 1st generation of chondrocytes in the proportion of 1:1 were mixed. The pellet at the final concentration of 1×10⁶/mL was set as the experimental group, and the considerable population of chondrocytes and synovial-derived mesenchymal stem cells was set as the control group. After cultivated in vitro for 21 days, the newly-born cartilage was evaluated through morphological observation, histological stain, and RT-PCR detection products. Results: After cultivated for 21 days, pellet chondroid tissue was formed and presented tenacious texture and milk white. Type-II collagen was identified by immunohistochemical staining. Matrix could be stained by Type-II collagen. RT-PCR results indicated that pellet expressed cartilage specific genes Type-II collagen and proteoglycan. Conclusion: Through direct co-culture, synovial-derived mesenchymal stem cells and chondrocytes can form mature cartilage.

中图分类号:

R329.2

邵博 ,胡露露 ,龚忠诚 ,杨萌 ,宁晓婷 ,王玥 ,胡鑫 ,刘慧
克热木·阿巴斯 ,凌彬 ,尹小朋 ,高芸竹 ,林兆全. 滑膜间充质干细胞与软骨细胞接触式共培养构建软骨的初步研究[J]. 口腔医学研究, 2016, 32(6): 554-558.

SHAO Bo, HU Lu-lu, GONG Zhong-cheng, YANG Meng, NING Xiao-ting, WANG Yue, HU Xin, LIU Hui, KEREMU·ABAShi, LIN Bin, YING Xiao-peng, GAO Yun-zu, LIN Zao-quan.. Direct Co-culture of Synovial Stem Cells and Chondrocytes Induces Enhanced Cartilage Tissue Formation.[J]. Journal of Oral Science Research, 2016, 32(6): 554-558.

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滑膜间充质干细胞与软骨细胞接触式共培养构建软骨的初步研究

Direct Co-culture of Synovial Stem Cells and Chondrocytes Induces Enhanced Cartilage Tissue Formation.

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