体外共培养诱导人羊膜上皮细胞向软骨细胞分化的初步研究  

Preliminary study on inducing differentiation of human amniotic epithelial cells into articular chondrocytes byco-culture invitro

作　　者：王康[1] 智晓东 张玉强 张文景 王伟 WANG Kang;ZHI Xiaodong;ZHANG Yuqiang;ZHANG Wenjing;WANG Wei(The Affiliated People's Hospital of Ningbo University,Ningbo 315040,Zhejiang,China)

机构地区：[1]宁波大学附属人民医院,宁波315040 [2]锦州医科大学附属第一医院

出　　处：《现代实用医学》2023年第6期719-723,共5页Modern Practical Medicine

基　　金：辽宁省科学技术基金(20180551216);锦州医科大学校企合作基金(2020002)。

摘　　要：目的探讨人羊膜上皮细胞(hAECs)和软骨细胞共培养,是否能把hAECs诱导分化为软骨细胞。方法采用酶消化法分离获得hAECs,采用形态学观察、免疫荧光和流式细胞仪鉴定;运用Transwell非接触共培养系统将hAECs和软骨细胞共培养21 d,诱导其分化;通过光镜观察、免疫组织化学染色和甲苯胺蓝检测诱导后hAECs形态改变、Ⅱ型胶原蛋白和聚集蛋白多糖的表达;Western-bolt和RT-qPCR检测分化细胞COL1、COL2、AGGRECAN和SOX9的表达。结果体外培养的hAECs和软骨细胞的形态相似,免疫组化和甲苯胺蓝染色检测诱导后hAECs中Ⅱ型胶原蛋白和聚集蛋白多糖呈阳性表达;Western-bolt和RT-qPCR检测分化细胞中COL1、COL2、AGGRECAN和SOX9高表达。结论hAECs在与软骨细胞共培养的诱导条件下,可以向软骨细胞分化。Objective To investigate the feasibility of inducing differentiation of human amniotic membrane cells into articular chondrocytes by co-culturing with articular chondrocytes.Methods Human amniotic epithelial cells were isolated by enzyme digestion and identified by morphological observation,immunofluorescence and flow cy-tometry.Human amniotic membrane cells and articular chondrocytes were co-cultured for 21 days with Transewell non-contact co-culture system to induce human amniotic membrane cells to differentiate.The morphological changes of human amniotic epithelial cells and the expression of type II collagen and aggregating protein polysaccharide were detected by light microscopy,immunohistochemical staining and toluidine blue.Western blot and RT-qPCR were used to detect the expression of COL1,COL2,AGGRECAN and SOX9 in differentiated cells.Results The morphology of human amniotic membrane cells cultured in vitro and articular chondrocytes was similar.The positive expression of type Ⅱ collagen and aggrecan polysaccharide in induced human amniotic membrane cells was detected by immu-nohistochemistry and toluidine blue staining.Western blot and RT-qPCR results showed that the differentiated cells expressed COLI,COL2,AGGRECAN and SOX9 significantly.Conclusions Human amniotic epithelial cells can be induced to differentiate into articular chondrocytes by co-culture with articular chondrocytes.

关 键 词：人羊膜上皮细胞 软骨细胞 分化 

分 类 号：R684.3[医药卫生—骨科学]