3种成软骨诱导培养基对单层培养和微球培养的人脂肪干细胞成软骨效果比较

doi:10.13418/j.issn.1001-165x.2021.01.014

中国临床解剖学杂志 ›› 2021, Vol. 39 ›› Issue (1): 71-75.doi: 10.13418/j.issn.1001-165x.2021.01.014

3种成软骨诱导培养基对单层培养和微球培养的人脂肪干细胞成软骨效果比较

于庆贺¹，　曲戎梅²，　张国炜¹，　李鑫¹，　仇显帅¹，　欧阳钧²，　戴景兴²，　闵少雄¹

1.南方医科大学珠江医院脊柱外科，广州 510280； 2.南方医科大学基础医学院解剖教研室暨
广东省医学生物力学重点实验，广州 510515

收稿日期:2019-09-27 出版日期:2021-01-25 发布日期:2021-01-27
通讯作者: 闵少雄，主任医师，E-mail：msxbear24@163.com 戴景兴，教授，E-mail：daijx@smu.edu.cn
作者简介:于庆贺（1992-），男，硕士研究生，研究方向：组织工程与再生医学，E-mail：442388223@qq.com
基金资助:
广东省自然科学基金（2017A030313564）；国家重点研发计划（2017YFC1105000）

Comparison of chondrogenic effects of three kinds of chondrogenic induction media on human adipose-derived stem cells in monolayer and microsphere cultured

Yu Qinghe¹, Qu Rongmei², Zhang Guowei¹, Li Xin¹, Qiu Xianshuai¹, Ouyang Jun², Dai Jingxing², Min Shaoxiong¹

1. Department of Spinal Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China; 2. Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Inspection in Tissue Engineering, Southern Medical University, Guangzhou 510515, China

Received:2019-09-27 Online:2021-01-25 Published:2021-01-27

摘要/Abstract

摘要： 目的　比较3种成软骨诱导培养基对单层培养和微球培养的人脂肪干细胞（human adipose mesenchymal stem cells，hASCs）体外成软骨分化的效果。方法　从人脂肪组织中提取hASCs，分别采用单层培养和微球培养，加入3种不同成分的成软骨诱导培养基，A组：胎牛血清（FBS）+ 10 ng/ml转化生长因子3（TGF-β3）；B组：胰岛素-转铁蛋白-亚硒酸钠（ITS）+ 1 ng/ml TGF-β3；C组：ITS+10 ng/mL TGF-β3。3周后提取单层培养的细胞蛋白进行成软骨相关标志物检测，并对细胞微球进行阿尔新蓝、甲苯胺蓝、天狼星红染色，比较成软骨效果。结果　3周后单层培养体系C组的软骨标志物表达最高，微球培养阿尔新蓝、甲苯胺蓝、天狼星红染色C组着色最深。结论　作为培养基添加物，ITS在诱导hASCs成软骨分化时的效果优于FBS，而作为刺激hASCs成软骨的重要生长因子TGF-β3，10 ng/ml的浓度比1 ng/ml作用更强。

关键词: 人脂肪干细胞（hASCs）, 　成软骨分化, 　胎牛血清（FBS）, 　转化生长因子3（TGF-β3）

Abstract: Objective　To compare the chondrogenic differentiation of human adipose mesenchymal stem cells (hASCs) in vitro by three components of chondrogenic induction medium.　Methods　hASCs were extracted from human adipose tissues and then performed in monolayer culture and microsphere culture respectively. Three kinds of different components of chondrogenic induction medium (group A: Fetal bovine serum (FBS) + high concentration TGF-β3, group B: Insulin, Transferrin, Selenium Solution (ITS) + low concentration TGF-β3, group C: ITS + high concentration TGF-β3) were added. After 3 weeks, the cell proteins in monolayer culture were extracted for detection of cartilage-related markers, and the cell microspheres were stained with Toluidine blue, Alcian blue, Sirius red to compare the effect of cartilage formation. Results 　After 3 weeks, the expression of cartilage markers was the highest in the group C of monolayer culture system, and the staining of microspheres was also the deepest in the group C.　Conclusions　As a medium additive, ITS has a better effect than FBS in inducing chondrogenic differentiation of hASCs. As an important growth factor stimulating chondrogenesis of hASCs, the concentration of 10 ng/ml TGF-β3 has a stronger effect than the concentration of 1 ng/ml.

Key words: 人脂肪干细胞（hASCs）, 　成软骨分化, 　胎牛血清（FBS）, 　转化生长因子3（TGF-β3）

中图分类号:

R684

于庆贺, 曲戎梅, 张国炜, 李鑫, 仇显帅, 欧阳钧, 戴景兴, 闵少雄. 3种成软骨诱导培养基对单层培养和微球培养的人脂肪干细胞成软骨效果比较[J]. 中国临床解剖学杂志, 2021, 39(1): 71-75.

Yu Qinghe, Qu Rongmei, Zhang Guowei, Li Xin, Qiu Xianshuai, Ouyang Jun, Dai Jingxing, Min Shaoxiong. Comparison of chondrogenic effects of three kinds of chondrogenic induction media on human adipose-derived stem cells in monolayer and microsphere cultured[J]. Chinese Journal of Clinical Anatomy, 2021, 39(1): 71-75.

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3种成软骨诱导培养基对单层培养和微球培养的人脂肪干细胞成软骨效果比较

Comparison of chondrogenic effects of three kinds of chondrogenic induction media on human adipose-derived stem cells in monolayer and microsphere cultured

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