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人脐带静脉间充质干细胞的表征与基因操作:在基于细胞的基因治疗中的潜在应用。

Characterization and genetic manipulation of human umbilical cord vein mesenchymal stem cells: potential application in cell-based gene therapy.

作者信息

Kermani Abbas Jafari, Fathi Fardin, Mowla Seyed Javad

机构信息

Department of Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran.

出版信息

Rejuvenation Res. 2008 Apr;11(2):379-86. doi: 10.1089/rej.2008.0674.

Abstract

Stem cells are defined by two main characteristics: self-renewal capacity and commitment to multi-lineage differentiation. The cells have a great therapeutic potential in repopulating damaged tissues as well as being genetically manipulated and used in cell-based gene therapy. Umbilical cord vein is a readily available and inexpensive source of stem cells that are capable of generating various cell types. Despite the recent isolation of human umbilical cord vein mesenchymal stem cells (UVMSC), the self-renewal capacity and the potential clinical application of the cells are not well known. In the present study, we have successfully isolated and cultured human UVMSCs. Our data further revealed that the isolated cells express the self-renewal genes Oct-4, Nanog, ZFX, Bmi-1, and Nucleostemin; but not Zic-3, Hoxb-4, TCL-1, Tbx-3 and Esrrb. In addition, our immunocytochemistry results revealed the expression of SSEA-4, but not SSEA-3, TRA-1-60, and TRA-1-81 embryonic stem cell surface markers in the cells. Also, we were able to transfect the cells with a reporter, enhanced green fluorescent protein (EGFP), and a therapeutic human brain-derived neurotrophic factor (hBDNF) gene by means of electroporation and obtained a stable cell line, which could constantly express both transgenes. The latter data provide further evidence on the usefulness of umbilical cord vein mesenchymal stem cells as a readily available source of stem cells, which could be genetically manipulated and used in cell-based gene therapy applications.

摘要

干细胞由两个主要特征定义

自我更新能力和向多谱系分化的潜能。这些细胞在修复受损组织以及进行基因操作并用于基于细胞的基因治疗方面具有巨大的治疗潜力。脐静脉是一种易于获取且成本低廉的干细胞来源,能够产生多种细胞类型。尽管最近已分离出人类脐静脉间充质干细胞(UVMSC),但其自我更新能力和潜在的临床应用尚不清楚。在本研究中,我们成功分离并培养了人类UVMSC。我们的数据进一步显示,分离出的细胞表达自我更新基因Oct-4、Nanog、ZFX、Bmi-1和Nucleostemin;但不表达Zic-3、Hoxb-4、TCL-1、Tbx-3和Esrrb。此外,我们的免疫细胞化学结果显示细胞中表达SSEA-4,但不表达胚胎干细胞表面标志物SSEA-3、TRA-1-60和TRA-1-81。而且,我们能够通过电穿孔用报告基因增强型绿色荧光蛋白(EGFP)和治疗性人类脑源性神经营养因子(hBDNF)基因转染细胞,并获得了能持续表达这两种转基因的稳定细胞系。后者的数据进一步证明了脐静脉间充质干细胞作为一种易于获取的干细胞来源的有用性,其可进行基因操作并用于基于细胞的基因治疗应用。

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