摘要
目的观测单克隆人永生化骨髓基质干细胞（hMSC－TERT）体内外分化潜能和自分泌血管内皮细胞生长因子（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。将各单克隆细胞卜清液中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、外分化能力存在差异。细胞十清液中，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．FERT20，7．684±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－TER3细胞进行临床和实验研究做了有益的探讨。
Objective To investigate the multi-differentiation potential and VEGF secretory, volume of monoclonal immortalized hmnan 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 adipoeytic, osteogenic, neuronal differentiation potential in vitro were determined by Oil Red O staining, Von Kossa staining and immuocytochemistry for Tubulin-β Ⅲ 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 difference between different nmoclunal 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 5^th day. The positive rates of CK in grafts formed by those monoclonal 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 monoclona/ hMSC-TERT may direct correlation with the epithelial differentiation potential of those cells.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2011年第17期1193-1196,共4页
National Medical Journal of China
基金
国家自然科学基金（30672156）
关键词
骨髓基质细胞
干细胞
分化
血管内皮细胞生长因子
上皮细胞
Mesenchymal stem cells
Stem cell
Differentiation
Vascular endothelial growth factor
Epithelial
