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首页> 外文期刊>Cell and Tissue Research >Xeno-free expansion of adult keratinocytes for clinical application: the use of human-derived feeder cells and serum
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Xeno-free expansion of adult keratinocytes for clinical application: the use of human-derived feeder cells and serum

机译：成人角质形成细胞的无异卵膨胀，用于临床应用：使用人源性饲养细胞和血清

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Cultured epithelial autograft (CEA) was the birth of skin tissue engineering and encompassed methodologies for the isolation and expansion of autologous basal keratinocytes for burn treatment that are still practiced at some specialised units around the world. One of the limitations of CEA, however, is the reliance on animal-derived material during the manufacturing process and despite all efforts to date, no xeno-free alternative with proven efficacy has been reported. Here, we investigate whether human-derived fibroblast feeder cells and human serum can sufficiently and effectively provide a suitable microenvironment for adult keratinocyte isolation and expansion. Human dermal fibroblasts and epidermal keratinocytes were isolated from discarded skin during abdominoplasty and breast reduction procedures and cultured in xeno-free conditions. We report that these xeno-free adult keratinocytes form similar numbers of colony-forming units as those cultured using the Green's methods; however, xeno-free keratinocytes express lower levels of 6 integrin (CD49f; a progenitor and stem cell marker). We identified IL-8 as a potential growth factor secreted by adult human fibroblasts that may enhance keratinocyte colony formation in human serum. Finally, we propose a step-by-step xeno-free isolation and cultivation methodology for adult keratinocytes that can be tested further in serial cultivation for clinical application.
机译：培养的上皮自体移植（CEA）是皮肤组织工程的诞生，并包含了用于燃烧治疗的自体基底角质形成细胞的分离和扩展的方法，这些方法在世界各地的一些专业单位仍然实行。然而，CEA的局限性之一是在制造过程中依赖于动物衍生材料，尽管迄今为止迄今为止，但据报道，迄今为止没有验证的疗效的无异形替代品。在这里，我们研究人源性的成纤维细胞和人血清是否可以充分且有效地为成人角质形成细胞分离和膨胀提供合适的微环境。在腹部成形术期间，在腹部成形术期和乳房减少手术期间分离出人的皮肤成纤维细胞和表皮角表纤维细胞，并在无异卵状条件下培养。我们报告说，这些无异种成人角质形成细胞形成了类似数量的菌落形成单元，因为使用绿色方法培养的单位;然而，无异叶角蛋白细胞表达较低水平的6整联蛋白（CD49F;祖细胞和干细胞标记物）。我们将IL-8鉴定为可通过成年人成纤维细胞分泌的潜在生长因子，其可以增强人血清中的角质形成细胞菌落形成。最后，我们为成人角质形成细胞提出了一步一步的异象隔离和培养方法，其可以在临床应用的连续培养中进一步测试。

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