CAJ | 학술논문

目的观测单克隆人永牛化骨髓基质干细胞(hMSC-TERT)体内外分化潜能和自分泌血管内皮细胞牛长因子(VEGF)的相关性.方法梯度稀释法建立单克隆hMSC-TERT,ELISA方法测定细胞培养液中VEGF含量.将各单克隆细胞进行脂肪、骨和神经样细胞体外诱导,检测多向分化能力.接种联合免疫缺陷性鼠(SCID),免疫组化法检测细胞体内分化率.将各单克隆细胞上清液中VEGF的含量变化值与其上皮细胞分化率进行单变量方差分析.结果各hMSC-TERT细胞的体内、外分化能力存在筹异.细胞上清液中,第5天VEGF浓度较第3天明显减少.各细胞上清液中VEGF浓度的减少幅度差异有统计学意义(hMSC-TERT2,22 ±0.33;hMSC-TERT20,7.68±0.25;hMSC-TERT-C2,103±35.67;hMSC-TERT-C3,140±34.17;hMSC-TERT-C6,115±28.75;hMSC-TERT-C13,-21±11;hMSC-TERT-C19,181±52.92;hMSC-TERT-C20,101±27.08;hMSC-TERT-C21,-29±10.5;hMSC-TERT-C29,46±25.42),减少幅度与细胞体内分化时角蛋白的阳性率呈正相关.结论单克隆hMSC-TERT上清液中VEGF变化幅度与细胞体上皮分化能力呈正相关.为选用特定的单克隆hMSC-TERT细胞进行临床和实验研究做了有益的探讨.
목적 관측단극륭인영우화골수기질간세포(hMSC-TERT)체내외분화잠능화자분비혈관내피세포우장인자(VEGF)적상관성.방법 제도희석법건립단극륭hMSC-TERT,ELISA방법측정세포배양액중VEGF함량.장각단극륭세포진행지방、골화신경양세포체외유도,검측다향분화능력.접충연합면역결함성서(SCID),면역조화법검측세포체내분화솔.장각단극륭세포상청액중VEGF적함량변화치여기상피세포분화솔진행단변량방차분석.결과 각hMSC-TERT세포적체내、외분화능력존재주이.세포상청액중,제5천VEGF농도교제3천명현감소.각세포상청액중VEGF농도적감소폭도차이유통계학의의(hMSC-TERT2,22 ±0.33;hMSC-TERT20,7.68±0.25;hMSC-TERT-C2,103±35.67;hMSC-TERT-C3,140±34.17;hMSC-TERT-C6,115±28.75;hMSC-TERT-C13,-21±11;hMSC-TERT-C19,181±52.92;hMSC-TERT-C20,101±27.08;hMSC-TERT-C21,-29±10.5;hMSC-TERT-C29,46±25.42),감소폭도여세포체내분화시각단백적양성솔정정상관.결론 단극륭hMSC-TERT상청액중VEGF변화폭도여세포체상피분화능력정정상관.위선용특정적단극륭hMSC-TERT세포진행림상화실험연구주료유익적탐토.
Objective To investigate the multi-differentiation potential and VEGF secretory volume of monoclonal immortalized human mesenchymal stem cells(hMSC-TERT)and to determine the relationship between them.Methods Monoelonal hMSC-TERT were isolated using limiting dilution.The growth curves of them were detected by method of MTT.ELISA was used to detect the concentration of VEGF in the supernatant of those mnoclonal hMSC-TERT.Their adipocytic,osteogenic,neuronal differentiation potential in vitro were determined by Oil Red O staining,Van Kossa staining and immuocytochemistry for Tubulin-βⅢ antibody.Those Monoclonal hMSC-TERT were transplanted into the subcutaneously of severe combined immunodeficiency(SCID) mices.The grafts of those cells were removed and analyzed by immunohistochemistry for pathologic tissue markers to discover the multi-differentiation potential of those mnoclonal hMSC-TERT in vivo.Results There was statistically significant difieFence between different mnoelonal hMSC-TERT in multidifferentiation potential and the decreased concentration of VEGF in the supernatant of those mnoconal hMSC-TERT from the 3 rd day to the 5th day.The positive rates of CK in grafts formed by those monoelonal hMSC-TERT in SCID mices were direct correlation with the decreased concentration of VEGF in the supernatant of those cells.Conclusion The secretory capability of VEGF of those monoclonal hMSC-TERT may direct correlation with the epithelial differentiation potential of those cells.