成人骨髓基质细胞体外培养及其生物学特性

摘要

Objective To explore in vitro culture method of human bone mesenchymal stem cells (BMSCs) of cerebral hemorrhage sequelae patients and its amplified biological characteristics.Methods Ilium myeloid tissues were taken from patients with 3-month cerebral hemorrhage sequelae, and they were in adherence standing culture after removing the lymphocyte, and multiple liquid purification. Morphology and growth characteristics were examined by inverted phase contrast microscope. Cell surface markers CD29, CD34, CD45, and CD105 in the third and sixth generation were tested by flow cytometer.Results After 48 h of phase contrast microscopy inoculate, there were several spindle-shaped cells. After 6~8 d, cells were in colony. After passage, cellular morphology tended to be conform and swirling. Flow cytometer showed that CD34 and CD45 were positive, while CD29 and CD105 were negative.Conclusion In vitro density gradient centrifugation and adherence standing method can provide a large number of homogeneous BMSCs.%目的：探索体外培养脑出血后遗症患者骨髓基质细胞(BMSCs)的方法及扩增后的生物学特性。方法取脑出血后3个月以上患者髂骨骨髓组织,去除淋巴细胞后采用静置贴壁培养法接种培养,经多次换液纯化。在倒置相差显微镜下观察细胞形态,取第3、6代应用流式细胞仪检测细胞表面抗原CD29、CD34、CD45、CD105因子的表达。结果倒置相差显微镜下接种48 h后,发现少数细胞贴壁,呈梭形,6~8 d后细胞形成集落。传代后,细胞形态趋于一致,可成漩涡状。流式细胞仪检测显示CD34、CD45阴性,而CD29、CD105阳性。结论体外密度梯度离心法和贴壁静置可以获得大量高度均一的表型符合骨髓基质细胞的细胞。