冻存人脐带间充质干细胞的成软骨分化  

Chondrogenic differentiation of cryopreserved human umbilical cord derived mesenchymal stem cells

作　　者：李星儿[1] 邓天明[1] 刘亭 黄桢钧 徐盎然[3] Li Xing’er;Deng Tianming;Liu Ting;Huang Zhenjun;Xu Ang’ran(Department of General Medicine,Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,China;Interdisciplinary Research Center of Frontier Medicine,Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,China;Department of Integrated Medicine,Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,China)

机构地区：[1]广州医科大学附属第五医院全科医学科,广州510700 [2]广州医科大学附属第五医院前沿医学交叉研究中心,广州510700 [3]广州医科大学附属第五医院综合病区,广州510700

出　　处：《中华生物医学工程杂志》2022年第5期502-507,共6页Chinese Journal of Biomedical Engineering

基　　金：广东省自然科学基金(2018A030310168);广州市属高校科研项目青年项目(1201630067)。

摘　　要：目的:研究从冻存人脐带组织中分离培养的脐带间充质干细胞(hUC-MSCs)的成软骨分化能力。方法:从冻存人脐带组织中分离培养的hUC-MSCs分为诱导组与对照组,分别采用成软骨诱导分化完全培养基和常规培养基培养7 d。采用实时荧光定量(qRT)-PCR法检测干细胞中成软骨标志物ColⅡa1、ColⅩa1和蛋白聚糖mRNA的表达,免疫印迹及免疫荧光染色检测干细胞Ⅱ型胶原蛋白的表达,阿尔新蓝染色检测干细胞中氨基聚糖含量。结果:hUC-MSCs成软骨诱导分化7 d后,与对照组比较,干细胞中ColⅡa1、ColⅩa1和蛋白聚糖mRNA的表达均上升,Ⅱ型胶原蛋白的表达上升,氨基聚糖含量升高(均 P<0.05)。 结论:从冻存人脐带组织中分离培养的hUC-MSCs采用成软骨诱导分化完全培养基培养7 d后可分化为成软骨细胞,可用于软骨疾病的实验研究和临床治疗。Objective To investigate the chondrogenic differentiation of umbilical cord derived mesenchymal stem cells isolated and cultured from cryopreserved human umbilical cord(hUC-MSCs).Methods hUC-MSCs isolated and cultured from cryopreserved human umbilical cord were divided into the induction group and control group,and were cultured in complete chondrogenic differentiation medium and conventional medium for 7 days,respectively.The expression of chondrogenic markers ColⅡa1,Col X a1 and Aggrecan mRNA in the hUC-MSCs was examined by real-time fluorescence quantitative(qRT)-PCR,and the expression of collagen typeⅡprotein in the hUC-MSCs was examined by immunoblotting analysis and immunofluorescence staining.The content of glycosaminoglycans(GAG)in the hUC-MSCs was examined by Alcian blue staining.Results After 7 days of chondrogenic differentiation,the hUC-MSCs showed higher mRNA expression of ColⅡa1,Col X a1 and Aggrecan,higher protein expression of collagen typeⅡ,and higher content of GAG,compared with the control group(all P<0.05).Conclusion hUC-MSCs isolated and cultured from cryopreserved human umbilical cord can differentiate into chondrocytes after 7-day induction in chondrogenic differentiation medium.This can be useful for experimental research and clinical treatment on cartilage diseases.

关 键 词：间质干细胞 软骨细胞 脐带 细胞分化 冷冻 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]