CAJ | 학술논문

背景:目前,国内外对骨髓间充质干细胞、胰腺干细胞向胰岛β样细胞分化的研究报道较多,但关于脐血间充质干细胞向胰岛β样细胞分化的研究报道较少.目的:验证脐血间充质干细胞能否在体外向胰岛β样细胞分化,并探索其诱导条件.方法:在无菌条件下采集正常产妇脐血,用羟乙基淀粉沉淀法分离脐血中的有核细胞,进而采用贴壁筛选法获得脐血间充质干细胞.纯化后的脐血间充质干细胞用表皮生长因子、β-巯基乙醇、高糖、激活素A和肝细胞生长因子进行诱导.观察诱导后的细胞形态变化,采用胰岛素免疫荧光染色对诱导后的细胞进行鉴定,定量检测胰岛素分泌水平及其对葡萄糖刺激的反应性.结果与结论:经过诱导后,细胞形态发生明显变化,形态变圆而且聚集成团;诱导后细胞的胰岛素免疫荧光染色为阳性;而且细胞能分泌少量胰岛素,并对糖刺激具有反应性.在体外,脐血间充质干细胞具有向胰岛β样细胞分化的潜能.
배경:목전,국내외대골수간충질간세포、이선간세포향이도β양세포분화적연구보도교다,단관우제혈간충질간세포향이도β양세포분화적연구보도교소.목적:험증제혈간충질간세포능부재체외향이도β양세포분화,병탐색기유도조건.방법:재무균조건하채집정상산부제혈,용간을기정분침정법분리제혈중적유핵세포,진이채용첩벽사선법획득제혈간충질간세포.순화후적제혈간충질간세포용표피생장인자、β-구기을순、고당、격활소A화간세포생장인자진행유도.관찰유도후적세포형태변화,채용이도소면역형광염색대유도후적세포진행감정,정량검측이도소분비수평급기대포도당자격적반응성.결과여결론:경과유도후,세포형태발생명현변화,형태변원이차취집성단;유도후세포적이도소면역형광염색위양성;이차세포능분비소량이도소,병대당자격구유반응성.재체외,제혈간충질간세포구유향이도β양세포분화적잠능.
BACKGROUND: At present,there are many reports regarding the differentiation of bone marrow-derived mesenchymal stem cells or pancreatic gland stem cells into pancreatic islet β-like cells.But little is about umbilical cord blood-derived mesenchymal stem cells(UCBMSCs)differentiation into pancreatic islet β-like cells in vitro.OBJECTIVE: To investigate whether UCBMSCs can differentiate into pancreatic islet β-like cells in vitro and the optimal inducing condition.METHODS: UCB samples were obtained sterilely from healthy parturients.Nucleated cells were isolated by sedimentation with hydroxyethyl starch and MSCs were obtained by adherent method.Then purified UCBMSCs were induced with epidermal growth factor,β-mercaptoethanol,high glucose,Activin A and hepatocyte growth factor(HGF).Following cell morphology observation,induced cells were identified by insulin immunofluorescence.In addition,insulin secretion and glucose-stimulated insulin release were examined by chemiluminescence immunoassay.RESULTS AND CONCLUSION: After induction,many cells exhibited a round appearance and produced islet-like cell clusters.Immunofluorescence assay showed insulin positive in the treated cells.In addition,chemiluminescence immunoassay demonstrated low expression of insulin and secretion of insulin upon glucose challenge.UCBMSCs can differentiate into pancreatic islet β-like cells in vitro.