CAJ | 학술논문

背景:人脐带沃顿胶间充质干细胞满足了国际细胞治疗协会规定的间充质干细胞的特点,能够向骨、软骨、脂肪、肌肉、神经细胞诱导分化并支持其他干细胞的扩增,对免疫系统有良好的耐受性,对肿瘤有定向迁徙性.目的:观察人脐带沃顿胶间充质干细胞与脑肿瘤干细胞体外共培养后,脑肿瘤干细胞的生物学变化.方法:以原位法培养人脐带沃顿胶间充质干细胞,以酶消化法培养人脑肿瘤组织脑肿瘤干细胞,以细胞传代法获取第3代细胞.应用不添加任何生长因子的无血清培养基将两种细胞在24孔板中进行直接共培养.第3,7天应用流式细胞术检测细胞CD133表达;应用免疫荧光法检测贴壁细胞巢蛋白和胶质纤维酸性蛋白表达;将第3天离心所得的共培养上清液重悬第3代脑肿瘤干细胞并与正常培养悬浮的第3代脑肿瘤干细胞置入96孔板中,应用酶标仪检测两组细胞生长曲线的差异.结果与结论:两种细胞共培养后在倒置显微镜下可见脑肿瘤干细胞球随着培养时间的增加出现分解、贴壁、分化现象;贴壁的脑肿瘤干细胞免疫荧光染色胶质纤维酸性蛋白和巢蛋白均阳性.恶性程度高的脑肿瘤组织培养的脑肿瘤干细胞表达CD133量越高,而与沃顿胶间充质干细胞共培养后随着时间的变化均出现CD133表达量降低.共培养3 d的上清液培养的脑肿瘤干细胞与正常培养基培养的脑肿瘤干细胞相比,增殖明显受到抑制.结果显示沃顿胶间充质干细胞与脑肿瘤干细胞体外共培养后可限制脑肿瘤干细胞表面标记物CD133的阳性率以及细胞增殖能力并促使其分化.
배경:인제대옥돈효간충질간세포만족료국제세포치료협회규정적간충질간세포적특점,능구향골、연골、지방、기육、신경세포유도분화병지지기타간세포적확증,대면역계통유량호적내수성,대종류유정향천사성.목적:관찰인제대옥돈효간충질간세포여뇌종류간세포체외공배양후,뇌종류간세포적생물학변화.방법:이원위법배양인제대옥돈효간충질간세포,이매소화법배양인뇌종류조직뇌종류간세포,이세포전대법획취제3대세포.응용불첨가임하생장인자적무혈청배양기장량충세포재24공판중진행직접공배양.제3,7천응용류식세포술검측세포CD133표체;응용면역형광법검측첩벽세포소단백화효질섬유산성단백표체;장제3천리심소득적공배양상청액중현제3대뇌종류간세포병여정상배양현부적제3대뇌종류간세포치입96공판중,응용매표의검측량조세포생장곡선적차이.결과여결론:량충세포공배양후재도치현미경하가견뇌종류간세포구수착배양시간적증가출현분해、첩벽、분화현상;첩벽적뇌종류간세포면역형광염색효질섬유산성단백화소단백균양성.악성정도고적뇌종류조직배양적뇌종류간세포표체CD133량월고,이여옥돈효간충질간세포공배양후수착시간적변화균출현CD133표체량강저.공배양3 d적상청액배양적뇌종류간세포여정상배양기배양적뇌종류간세포상비,증식명현수도억제.결과현시옥돈효간충질간세포여뇌종류간세포체외공배양후가한제뇌종류간세포표면표기물CD133적양성솔이급세포증식능력병촉사기분화.
BACKGROUND:Human mesenchymal stem cells derived from Wharton's jelly(WJCs)display the characteristics of MSCs as defined by the International Society for Cellular Therapy.They can be differentiated into bone,cartilage,adipose,muscle,and neural cells.They can also support the expansion of other stem cells,be weli-tolerated by the immune system,and have the ability to home to tumors.OBJECTIVE:To investigate biological changes of WJCs and brain tumor stem cells(BTSCs)co-cultured in vitro.METHODS:WJCs cultured by situ cultivation and BTSCs used enzyme digestion way respectively,and gathering the 3rd passage of WJCs though subculturing as well as BTSCs.Two kinds of cells co-cultured in 24-well plates in serum-free medium (SFM)without any growth factor.3 and 7 days after co-cultured respectively,CD133 expression of suspension cells in the 24-well plates were identified by flow cytometry,and immunofluorescence was performed for Nestin and glial fibrillary acidic protein (GFAP)expression of adherent cells.Co-culture supernatant(CCS)re-suspended 3~(rd) passage of BTSCs and cultured into 96-well plates at day 3,which were used to determine the difference in cell growth curve in both groups using a microplate reader.RESULTS AND CONCLUSION:With the cocultivation days increasing,the phenomenon that tumor sphere cells began to be decomposed,adherent and differentiated observed by an inverted microscope.BTSCs in the co-cultured group expressed GFAP and Nestin when adherent and differentiated.The higher degree of malignant brain tumor tissue used in culturing BTSCs was,the higher expression of CD133 in BTSCs was.CD 133~+ in BTSCs declined when co-cultured with WJCs.Growth curve of brain tumor stem cells cultured in CCS compared with in SFM at day 3,which indicates that the proliferation of BTSCs inhibited obviously.Results indicated that CD 133~+ expression and proliferative capacity of BTSCs went down and BTSCs underwent differentiation during the co-culture in vitro.