人胎盘不同组织分离间充质干细胞的生物学特性

人胎盘不同组织分离间充质干细胞的生物学特性 洪 艳，霍思维，陆 瑶，章 毅 上海市脐带血造血干细胞库，中国干细胞集团上海生物科技有限公司，上海市 201815 Biological characters of mesenchymal stem cells separated from different components of human placenta Hong Yan, Huo Si-wei, Lu Yao, Zhang Yi Shanghai Cord Blood Bank, Shanghai Stem Cell Technology Co., Ltd., Shanghai 201815, China 摘要 背景:胎盘间充质干细胞来源稳定，正逐渐成为广受关注的再生医学种子细胞来源。 目的:比较从胎盘组织绒毛膜与绒毛滋养层分离获取人胎盘间充质干细胞生物学特性的差异。 方法:手术剪剥离人胎盘面羊膜，分别剪取胎儿侧的绒毛膜与绒毛滋养层组织机械破碎后，分别加含?型胶原酶的PBS消 化液消化，分离为单个核细胞，用含有体积分数10%胎牛血清的DMEM培养基在37 ?、体积分数5%CO2、95%饱和湿 度下培养，48 h后全量换液，去除非贴壁细胞，添加新鲜培养基，当细胞融合90%左右时，胰酶传代。通过观察原代细胞总 数，细胞生长形态，以及间充质干细胞表面标记CD90、CD73、CD105的流式细胞测定结果比较从不同胎盘组织分离获得 胎盘间充质干细胞的差异。 结果与结论:流式细胞仪测定结果显示，从绒毛膜与绒毛滋养层中分离得到细胞间充质干细胞表面标记CD90、CD73以及 CD105的阳性率都在90%以上，两种来源的细胞生长都呈现典型的成纤维细胞形态，这表明绒毛膜与绒毛滋养层中分离得到 细胞都具有间充质干细胞的特性。提示消化时间相同，酶浓度相同，以及摇床转速相同的情况下，可从绒毛膜中可以得到更多 的细胞，对于后续培养更易获得较多的细胞。 中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血 干细胞;干细胞诱导;干细胞分化;组织工程 链接: 关键词 : 干细胞; 培养; 胎盘; 间充质干细胞; 绒毛膜; 绒毛滋养层; ?型胶原酶; 流式细胞仪; CD90; CD73; CD105; Abstract: BACKGROUND: Human placenta is a stable source for mesenchymal stem cells, which is becoming a cell source in the regenerative medicine that attracts widespread attentions. OBJECTIVE: To compare the biological characters of mesenchymal stem cells that separated from different components of human placenta (human chorion and villous trophoblast). METHODS: The amniotic membrane of placenta surface was detached surgically. Human chorion and villous trophoblast in the fetal side was cut into pieces. After digestion with PBS containing collagenase II, mononuclear cells were separated and cultured in Dulbecco’s modified Eagle’s medium containing 10% fetal bovine serum in 37 ? and 5% CO2, 95% saturated humidity, after 48 hours the full amount in liquid, dislodge suspension cells. Forty-eight hours later, the medium was changed completed, and non-adherent cells were removed. When cell fusion reached about 90%, trypsin digestion was employed for cell passage. Biological characters of mesenchymal stem cells separated from different components of human placenta were compared through observation of total number of primary cells, cell morphology, and surface markers expression (CD90, CD73 and CD105). RESULTS AND CONCLUSION: The flow cytometric analysis revealed that the cells separated from the human chorion and villous trophoblast were over 90% strongly positive for CD90, CD73, CD105. These two sources of cells showed typical fibroblast morphology, suggesting that the cells have the characteristics of mesenchymal stem cells. Under the same enzyme digestion time, enzyme concentration, and shaking speed, more cells are visible from the chorion, and the subsequent culture is easier to harvest cells. 中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血 干细胞;干细胞诱导;干细胞分化;组织工程 全文链接: Key words: stem cells; placenta; mesenchymal stem cells; chorion; collagenases; 中图分类号: R394.2 作者简介: 洪艳，女，1984年生，安徽省怀宁县人，汉族，2008年华东理工大学毕业，硕士，主要从事微生物学、干细胞、 细胞生物学、组织工程方面的研究。 引用本文: 洪 艳,霍思维,陆 瑶等. 人胎盘不同组织分离间充质干细胞的生物学特性[J]. 中国组织工程研究, 2014, 18(19): 3082-3087. Hong Yan,Huo Si-wei,Lu Yao et al. Biological characters of mesenchymal stem cells separated from different components of human placenta[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(19): 3082-3087.