Perin L, Giuliani S, Jin D, Sedrakyan S, Carraro G, Habibian R, Warburton D, Atala A, De Filippo R E
Childrens Hospital Los Angeles, Saban Research Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90027, USA.
Cell Prolif. 2007 Dec;40(6):936-48. doi: 10.1111/j.1365-2184.2007.00478.x.
The role of stem cells in regenerative medicine is evolving rapidly. Here, we describe the application, for kidney regeneration, of a novel non-genetically modified stem cell, derived from human amniotic fluid. We show that these pluripotent cells can develop and differentiate into de novo kidney structures during organogenesis in vitro.
Human amniotic fluid-derived stem cells (hAFSCs) were isolated from human male amniotic fluid obtained between 12 and 18 weeks gestation. Green fluorescent protein and Lac-Z-transfected hAFSCs were microinjected into murine embryonic kidneys (12.5-18 days gestation) and were maintained in a special co-culture system in vitro for 10 days. Techniques of live microscopy, histology, chromogenic in situ hybridization and reverse transcriptase polymerase chain reaction were used to characterize the hAFSCs during their integration and differentiation in concert with the growing organ.
Green fluorescent protein and Lac-Z-transfected hAFSCs demonstrated long-term viability in organ culture. Histological analysis of injected kidneys revealed that hAFSCs were capable of contributing to the development of primordial kidney structures including renal vesicle, C- and S-shaped bodies. Reverse transcriptase polymerase chain reaction confirmed expression of early kidney markers for: zona occludens-1, glial-derived neurotrophic factor and claudin.
Human amniotic fluid-derived stem cells may represent a potentially limitless source of ethically neutral, unmodified pluripotential cells for kidney regeneration.
干细胞在再生医学中的作用正在迅速演变。在此，我们描述了一种源自人羊水的新型非转基因干细胞在肾脏再生中的应用。我们表明，这些多能细胞在体外器官发生过程中可发育并分化为全新的肾脏结构。
从妊娠12至18周获取的男性人羊水中分离出人羊水来源的干细胞（hAFSCs）。将绿色荧光蛋白和Lac-Z转染的hAFSCs显微注射到妊娠12.5至18天的小鼠胚胎肾脏中，并在特殊的共培养系统中于体外维持10天。运用活细胞显微镜检查、组织学、显色原位杂交和逆转录聚合酶链反应技术，在hAFSCs与生长中的器官整合及分化过程中对其进行表征。
绿色荧光蛋白和Lac-Z转染的hAFSCs在器官培养中显示出长期活力。对注射后的肾脏进行组织学分析发现，hAFSCs能够促进包括肾小囊、C形和S形体在内的原始肾脏结构的发育。逆转录聚合酶链反应证实了早期肾脏标志物紧密连接蛋白-1、胶质细胞源性神经营养因子和闭合蛋白的表达。
人羊水来源的干细胞可能代表了一种用于肾脏再生的、伦理上中立且未经修饰的多能细胞的潜在无限来源。
