Qi Xiao-yuan, Sun Ze-lin, Liu Fang-jun, Li Chu-zhong, Zhang Ya-zhuo
Hebei United University Affiliated Hospital, Tangshan, Hebei Province 063000, China.
Zhonghua Yi Xue Za Zhi. 2011 May 10;91(17):1193-6.
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.
Monoclonal 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 monoclonal hMSC-TERT. Their adipocytic, osteogenic, neuronal differentiation potential in vitro were determined by Oil Red O staining, Von Kossa staining and immunocytochemistry for Tubulin-βIII antibody. Those Monoclonal hMSC-TERT were transplanted into the subcutaneously of severe combined immunodeficiency (SCID) mice. The grafts of those cells were removed and analyzed by immunohistochemistry for pathologic tissue markers to discover the multi-differentiation potential of those monoclonal hMSC-TERT in vivo.
There was statistically significant difference between different monoclonal hMSC-TERT in mult differentiation potential and the decreased concentration of VEGF in the supernatant of those monoclonal 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 mice were direct correlation with the decreased concentration of VEGF in the supernatant of those cells.
The secretory capability of VEGF of those monoclonal hMSC-TERT may direct correlation with the epithelial differentiation potential of those cells.
研究单克隆永生化人骨髓间充质干细胞（hMSC-TERT）的多向分化潜能及血管内皮生长因子（VEGF）分泌量，并确定两者之间的关系。
采用有限稀释法分离单克隆hMSC-TERT。用MTT法检测其生长曲线。用ELISA法检测单克隆hMSC-TERT上清液中VEGF的浓度。通过油红O染色、冯库萨染色及抗βIII微管蛋白抗体免疫细胞化学法测定其体外成脂、成骨、神经元分化潜能。将单克隆hMSC-TERT皮下移植到严重联合免疫缺陷（SCID）小鼠体内。取出细胞移植物，通过免疫组织化学分析病理组织标志物，以发现单克隆hMSC-TERT在体内的多向分化潜能。
不同单克隆hMSC-TERT在多向分化潜能上存在统计学显著差异，且这些单克隆hMSC-TERT上清液中VEGF浓度在第3天至第5天降低。SCID小鼠中由这些单克隆hMSC-TERT形成的移植物中细胞角蛋白（CK）阳性率与这些细胞上清液中VEGF浓度降低直接相关。
这些单克隆hMSC-TERT的VEGF分泌能力可能与这些细胞的上皮分化潜能直接相关。
