Selective ablation of human embryonic stem cells expressing a "suicide" gene
Selective ablation of human embryonic stem cells expressing a "suicide" gene
复制标题
DOI：
10.1634/stemcells.21-3-257
复制
发表时间：
2003-01-01
期刊：
影响因子：
5.2
通讯作者：
Benvenisty, N
中科院分区：
文献类型：
--
作者：
Schuldiner, M;Itskovitz-Eldor, J;Benvenisty, N
在过去的几年中，已经开发出了在移植医学中利用干细胞的技术程序。人胚胎干（ES）细胞可以产生无限数量的分化细胞，因此被认为是用于移植医学的细胞材料的潜在来源。然而，当移植后发生不受控制的细胞增殖时，可能会出现严重的临床问题。为了避免这些潜在问题，我们对人类 ES 细胞系进行了基因改造，使其表达单纯疱疹病毒胸苷激酶 (HSV-tk) 基因。 HSV-tk 蛋白的表达使 ES 细胞对美国食品和药物管理局批准的药物更昔洛韦敏感，在对其他细胞类型不致命的更昔洛韦浓度下诱导 HSV-tk(+) 细胞被破坏。即使在更昔洛韦的长期选择下，工程细胞的回复率也很低。 HSV-tk(+) 克隆保留了正常的核型和分化为来自所有三个胚层的细胞的能力。最重要的是，皮下注射 HSVtk(+) 人类 ES 细胞后小鼠体内出现的肿瘤可以通过给予更昔洛韦在体内消融。通过利用这些细胞系，可以提高涉及人类 ES 细胞来源的组织移植的安全水平。
Over the past few years, technological procedures have been developed for utilizing stem cells in transplantation medicine. Human embryonic stem (ES) cells can produce an unlimited number of differentiated cells and are, therefore, considered a potential source of cellular material for use in transplantation medicine. However, serious clinical problems can arise when uncontrolled cell proliferation occurs following transplantation. To avoid these potential problems, we genetically engineered human ES cell lines to express the herpes simplex virus thymidine kinase (HSV-tk) gene. Expression of the HSV-tk protein renders the ES cells sensitive to the U.S. Food and Drug Administration-approved drug ganciclovir, inducing destruction of HSV-tk(+) cells at ganciclovir concentrations that are nonlethal to other cell types. The reversion rate of engineered cells was low even under prolonged selection with ganciclovir. The HSV-tk(+) clones retained a normal karyotype and the ability to differentiate to cells from all three germ layers. Most importantly, tumors that arose in mice following subcutaneous injection of HSVtk(+) human ES cells could be ablated in vivo by administration of ganciclovir. By utilizing these cell lines, safety levels can be improved in transplantations involving tissues derived from human ES cells.