Human keratinocytes and derived products are crucial for skin repair and regeneration. Despite substantial advances in engineered skin equivalents, their poor availability and immunorejection remain major challenges in skin grafting. Induced keratinocyte-like cells (iKCs) were directly reprogrammed from human urine cells by retroviral transduction of two lineage-specific transcription factors BMI1 and △NP63α (BN). Expression of keratinocyte stem cell or their differentiation markers were assessed by PCR, immunofluorescence and RNA-Sequencing. Regeneration capacity of iKCs were assessed by reconstitution of a human skin equivalent under air-interface condition. BN-driven iKCs were similar to primary keratinocytes (pKCs) in terms of their morphology, protein expression, differentiation potential, and global gene expression. Moreover, BN-iKCs self-assembled to form stratified skin equivalents in vitro. This study demonstrated an approach to generate human iKCs that could be directly reprogrammed from human somatic cells and extensively expanded in serum- and feeder cell-free systems, which will facilitate their broad applicability in an efficient and patient-specific manner.

中文翻译：

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直接重编程的角质形成细胞样细胞的长期扩增和人皮肤的体外重建

人角质形成细胞及其衍生产品对于皮肤修复和再生至关重要。尽管工程皮肤等效物取得了重大进展，但它们的利用率差和免疫排斥仍然是皮肤移植的主要挑战。通过两个谱系特异性转录因子BMI1和△NP63α（BN）的逆转录病毒转导，可直接从人尿细胞中重新编程诱导的角质形成细胞样细胞（iKC）。通过PCR，免疫荧光和RNA测序评估角质形成细胞干细胞或其分化标志物的表达。iKC的再生能力是通过在空气界面条件下重建人体皮肤等效物来评估的。BN驱动的iKC在形态，蛋白质表达，分化潜能和整体基因表达方面与原代角质形成细胞（pKC）相似。此外，BN-iKC在体外自我组装形成分层的皮肤等效物。这项研究证明了一种生成人iKC的方法，该方法可以直接从人体细胞重新编程，并在无血清和饲养细胞的系统中广泛扩展，这将以有效且针对患者的方式促进其广泛应用。