Laboratoire de Génomique et Radiobiologie de la Kératinopoïèse, CEA/DRF/IBFJ/IRCM, Paris, France.
INSERM U967, Paris, France.
Methods Mol Biol. 2020;2109:155-167. doi: 10.1007/7651_2019_250.
The functional definition of somatic adult stem cells is based on their regenerative capacity, which allows tissue regeneration throughout life. Thus, refining methodologies to characterize this capacity is of great importance for progress in the fundamental knowledge of specific keratinocyte subpopulations but also for preclinical and clinical research, considering the high potential of keratinocytes in cell therapy. We present here a methodology which we define as iterative xenografting, which originates in the classical model of human skin substitute xenografts onto immunodeficient recipient mice. The principle of this functional assay is first to perform primary xenografts to assess graft take and the quality of epidermal differentiation. Then, human keratinocytes are extracted from primary graft samples to perform secondary xenografts, to assess the presence and preservation of functional keratinocyte stem cells with long-term regenerative potential. In the example of experiments shown, iterative skin xenografting was used to document the high regenerative potential of epidermal holoclone keratinocytes.
体干细胞的功能定义基于其再生能力，这种能力使组织在整个生命周期中得以再生。因此，改进用于鉴定这种能力的方法对于特定角质形成细胞亚群的基础知识的进展非常重要，同时对于临床前和临床研究也很重要，因为角质形成细胞在细胞治疗中有很高的潜力。我们在这里提出了一种我们定义为迭代异种移植的方法，它源于将人皮肤替代物异种移植到免疫缺陷受体小鼠上的经典模型。该功能测定的原理首先是进行原发性异种移植，以评估移植物的接受情况和表皮分化的质量。然后，从原发性移植物样本中提取人角质形成细胞，进行二次异种移植，以评估具有长期再生潜力的功能性角质形成细胞干细胞的存在和保存情况。在所示实验的例子中，迭代皮肤异种移植被用于证明表皮全克隆角质形成细胞的高再生潜力。
