人脐带血间充质干细胞条件培养基的制备及其生物学特性

Preparation and Biological Characteristics of Human Umbilical Cord Blood Mesenchymal Stem Cell Conditioned Medium

目的探讨人脐带血(human umbilical cord blood,hUCB)来源间充质干细胞(mesenchymal stem cells,MSCs)条件培养基(conditionedmedium,CM)的制备方法,并分析MSCs的旁分泌作用。方法无菌条件下收集足月健康新生儿的脐带血,经肝素抗凝,离心后分离、培养、传代hUCB-MSCs,用倒置显微镜观察不同时期细胞的生长情况和形态特征,并用酶标仪检测细胞增殖情况,待细胞融合达80%时更换培养基,再继续培养24h后收集培养上清液即为人脐带血间充质干细胞条件培养基(hUCB-MSCs-CM)。结果经过传代后,人脐带血间充质干细胞形态趋于均一,为梭形或成纤维样、体积较大、单个核,呈漩涡样生长,且增殖能力较强,能较好的制备出其条件培养基。结论能通过此方法成功制备人脐带血间充质干细胞条件培养基,其中含有MSCs旁分泌的各种生物活性因子,可以作为一种新的治疗来源应用于临床。

Objective Discussion on human umbilical cord blood（hUCB）-derived mesenchymal stem cells（MSCs） conditioned medium（CM） of the preparation method,and analyses the paracrine action of MSCs.Method Collection of neonatal cord blood under sterile conditions,the anticoagulant heparin,after centrifugation separation,culture,passage of hUCB-MSCs,to observe the growth and morphological characteristics of different periods of cells by inverted microscope,And with the enzyme labeled cell proliferation detection instrument for cell fusion,up to 80% when changing the culture medium,and then continue to develop after 24 hours of culture supernatant was collected for human umbilical cord blood mesenchymal stem cell conditioned medium（hUCB-MSCs-CM）.Results After passage,human umbilical cord blood mesenchymal stem cells tend to be homogeneous,fusiform or fibroblast-like,large volume,mononuclear,swirl like growth,and proliferation ability,can better prepare its conditioned medium.Conclusion Preparation of human umbilical cord blood mesenchymal stem cell conditioned medium by this method successfully,which contains secreting various bioactive factor MSCs,can be used as a new source of the therapy in clinical application.