Han Li-ying, Li Ya-ping, Ye Ming-zhu, Wang Bo-wei, Wang Qiang, Zhao Shu-hua, Li He-lian
Department of Obstetrics & Gynecology, Second Hospital, Jilin University, Changchun 130041, China.
Zhonghua Yi Xue Za Zhi. 2012 Nov 6;92(41):2930-3.
To explore the feasibility and safety of mdr1 gene transferred into placenta derived mesenchymal stem cells (P-MSCs) by reconstructed retroviral vector.
Human P-MSCs were isolated and expanded by Percoll density gradient and then transduced repeatedly by reconstructed retroviral vector containing mdr1 gene. The transfection and expression of mdr1 gene was detected by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS). Meanwhile, the biological features of mdr1-MSCs were identified and analyzed.
The expression of mdr1mRNA was found in transfected cells. The expression of P-glycoprotein (P-gp) encoded by mdr1 gene was (27.6 ± 5.1)% in the transfected P-MSCs cells versus (0.4 ± 0.1)% in the non-transfected P-MSCs cells (t = 14.291, P < 0.01). The percent of P-MSCs at quiescent phase (G0/G1 phase) was around 95.40% and it was in accord with the characterization of stem cells. The mdr1-MSCs exhibited typical ultrastructures of low-differentiated stem cells. Moreover, they still retained the potency of adipogenic and osteogenic differentiation in the presence of appropriate conditioned media.
A stable expression of P-gp may be obtained by reconstructed retroviral-mediated transfection in vitro. And transfected MSCs retain the characteristics of stem cells.
探讨用重组逆转录病毒载体将多药耐药1（mdr1）基因导入胎盘源间充质干细胞（P-MSCs）的可行性和安全性。
采用Percoll密度梯度法分离并扩增人P-MSCs，然后用含mdr1基因的重组逆转录病毒载体反复转导。通过逆转录-聚合酶链反应（RT-PCR）和荧光激活细胞分选（FACS）检测mdr1基因的转染和表达。同时，对mdr1-MSCs的生物学特性进行鉴定和分析。
在转染细胞中发现了mdr1mRNA的表达。mdr1基因编码的P-糖蛋白（P-gp）在转染的P-MSCs细胞中的表达为（27.6±5.1）%，而在未转染的P-MSCs细胞中为（0.4±0.1）%（t = 14.291，P < 0.01）。P-MSCs处于静止期（G0/G1期）的比例约为95.40%，这与干细胞的特征相符。mdr1-MSCs表现出低分化干细胞的典型超微结构。此外，在适当的条件培养基存在下，它们仍保留脂肪生成和成骨分化的潜能。
通过重组逆转录病毒介导的体外转染可获得P-gp的稳定表达。且转染后的间充质干细胞保留了干细胞的特性。
