Han Li-Ying, Ye Ming-Zhu, Li He-Lian, Wang Bo-Wei, Wang Qiang
Department of Obstetrics and Gynecology, Second Hospital, Jilin University, Changchun 130041, China.
Zhonghua Yi Xue Za Zhi. 2009 Oct 27;89(39):2793-6.
To transfer multidrug resistance gene (mdr1) into human placental mesenchymal stem cells (P-MSCs) by retroviral vector and assess the effects of mdr1 gene transduction upon biological features of P-MSCs.
Human P-MSCs were isolated from trypsin-digested term placentas and then transduced by reconstructed retroviral vector containing mdr1 gene and green fluorescent protein (GFP) reporter gene. Flow cytometric analysis was employed to determine the immunophenotypes of transfected P-MSCs. And the proliferation and cell cycle were detected by methyl thiazolyl tetrazolium and propidium iodide staining. Ultrastructures of transfected P-MSCs were observed and different induction conditions used to direct the cells to differentiate into adipocytes and osteoblasts.
The transfected P-MSCs still expressed stem cell markers such as CD29, CD44 and CD73. The mean cumulative time of population doubling was 23.9 hours. The cellular cycle retained the proliferative characterization of stem cells. Ultrastructural features of transfected P-MSCs included increased surface microvilli, abundant mitochondria and slightly swollen rough endoplasmic reticulum. Furthermore these transfected cells demonstrated osteogenic and adipogenic differentiation potentials under appropriate conditions.
The mdr1 gene transduction by retroviral vector in vitro has no significant effect upon biological characteristics of P-MSCs. It might provide experimental references for the application of P-MSCs in high-dose tumor chemotherapy.
通过逆转录病毒载体将多药耐药基因（mdr1）导入人胎盘间充质干细胞（P-MSCs），并评估mdr1基因转导对P-MSCs生物学特性的影响。
从胰蛋白酶消化的足月胎盘中分离出人P-MSCs，然后用含有mdr1基因和绿色荧光蛋白（GFP）报告基因的重组逆转录病毒载体进行转导。采用流式细胞术分析来确定转染后P-MSCs的免疫表型。并用甲基噻唑基四氮唑和碘化丙啶染色检测细胞增殖和细胞周期。观察转染后P-MSCs的超微结构，并使用不同的诱导条件将细胞诱导分化为脂肪细胞和成骨细胞。
转染后的P-MSCs仍表达干细胞标志物，如CD29、CD44和CD73。群体倍增的平均累积时间为23.9小时。细胞周期保留了干细胞的增殖特征。转染后P-MSCs的超微结构特征包括表面微绒毛增加、线粒体丰富和粗面内质网轻度肿胀。此外，这些转染细胞在适当条件下表现出成骨和成脂分化潜能。
体外逆转录病毒载体介导的mdr1基因转导对P-MSCs的生物学特性无显著影响。这可能为P-MSCs在大剂量肿瘤化疗中的应用提供实验参考。
