| Ad-hOSM转基因饲养层细胞对脐血CD34~+造血干/祖细胞增殖及归巢能力的影响 |
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| 引用本文： | 田丽娜,郁心,包婉蓉,陈瑶,韩亚丽,张凤娟,缪竟诚. Ad-hOSM转基因饲养层细胞对脐血CD34~+造血干/祖细胞增殖及归巢能力的影响[J]. 细胞与分子免疫学杂志, 2011, 27(3): 270-273 |
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| 作者姓名： | 田丽娜 郁心 包婉蓉 陈瑶 韩亚丽 张凤娟 缪竟诚 |
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| 作者单位： | 1. 苏州大学医学部细胞与分子生物学教研室,江苏,苏州,215123
2. 无锡市中心血站,江苏,无锡,214021 |
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| 基金项目： | 国防科工委基础科研计划资助项目(K0102061501) |
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| 摘 要： | 目的:建立转人抑瘤素M(hOSM)基因腺病毒载体的饲养层细胞,观察转基因细胞对脐血CD34+造血干/祖细胞扩增的影响,比较扩增前、后造血干/祖细胞体外迁移能力的变化。方法:建立转hOSM基因腺病毒载体的饲养层细胞,并用RT-PCR法和ELISA法鉴定目的基因;采用免疫磁珠法分离脐带血CD34+造血干/祖细胞,流式细胞术(FCM)检测纯度;将CD34+造血干/祖细胞与饲养层细胞共培养,FCM检测各组增殖效果;扩增后的造血干/祖细胞用跨膜迁移实验(Transwell实验)检测自发迁移率和SDF-1诱导迁移率以鉴定体外扩增的造血干/祖细胞的归巢能力。结果:建立的转基因饲养层细胞均有绿色荧光,RT-PCR法和ELISA法证实有目的基因表达,免疫磁珠法分离的脐血CD34+造血干/祖细胞纯度可达(96.8±2.28)%,与Ad-hOSM转基因饲养层细胞共培养7 d后CD34+造血干/祖细胞可扩增15.73倍,表面黏附分子CXCR4和CD54表达量仍较高,培养后的细胞用Tran-swell板做体外迁移实验,与转基因饲养层细胞共培养的干细胞,其诱导迁移率为(40.68±1.35)%,明显高于对照组,可以较好的保持其归巢能力。结论:转hOSM基因腺病毒载体的饲养层细胞可有效扩增脐血CD34+造血干/祖细胞,延缓其分化,并且体外扩增后仍保持较高的归巢能力。
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| 关 键 词： | hOSM 造血干/祖细胞 腺病毒载体 迁移率 |
The effect of transgenic cell strains expressing recombinant adenovirus vector of hOSM gene on the proliferation and homing ability of umbilical cord blood CD34+ hematopoietic stem/progenitor cells |
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| TIAN Li-na,YU Xin,BAO Wan-rong,CHEN Yao,HAN Ya-li,ZHANG Feng-juan,MIAO Jing-cheng. The effect of transgenic cell strains expressing recombinant adenovirus vector of hOSM gene on the proliferation and homing ability of umbilical cord blood CD34+ hematopoietic stem/progenitor cells[J]. Chinese journal of cellular and molecular immunology, 2011, 27(3): 270-273 |
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| Authors: | TIAN Li-na YU Xin BAO Wan-rong CHEN Yao HAN Ya-li ZHANG Feng-juan MIAO Jing-cheng |
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| Affiliation: | TIAN Li-na1,YU Xin2,BAO Wan-rong1,CHEN Yao1,HAN Ya-li1,ZHANG Feng-juan1,MIAO Jing-cheng1 1Cell and Molecular Biology Institute,College of Medicine,Soochow University,Suzhou 215123,2Wuxi Central Blood Station,Wuxi 214021,China |
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| Abstract: | AIM: To establish the transgenic cell strains expressing recombinant adenovirus vector of human Oncostain M(hOSM)gene which is supposed to be used as feeder layer cells for the proliferation of umbilical cord blood CD34+ hematopoietic stem/progenitor cell(HSPC) and compare its migration capacity before and after amplification in vitro.METHODS: Establish the transgenic cell strains expressing recombinant adenovirus vector of hOSM gene,and the objective gene was detected by RT-PCR and ELISA.The purity of umbi... |
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| Keywords: | human Oncostain M hematopoietic stem/progenitor cell(HSPC) adenovirus vector migration rate |
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