Chimeric composite skin substitutes for delivery of autologous keratinocytes to promote tissue regeneration.
Chimeric composite skin substitutes for delivery of autologous keratinocytes to promote tissue regeneration.
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DOI：
10.1097/sla.0b013e3181c1ab5f
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发表时间：
2010-02
影响因子：
9
通讯作者：
Allen-Hoffmann BL
中科院分区：
文献类型：
--
作者：
Rasmussen CA;Gibson AL;Schlosser SJ;Schurr MJ;Allen-Hoffmann BL
我们假设无病原体的人角质形成细胞祖细胞系NIKS以嵌合方式与患者的原代角质形成细胞培养，将产生完全分层的工程化皮肤替代组织，并用于将自体角质形成细胞递送至皮肤伤口。嵌合自体/同种异体生物工程皮肤替代品提供了一种创新的再生医学方法，用于提供伤口覆盖和恢复皮肤屏障功能，同时将自体角质形成细胞递送到伤口部位。NIKS角质形成细胞是用于该应用的有吸引力的同种异体细胞来源。使用GFP标记的NIKS和未标记的原代角质形成细胞的混合群体来模拟嵌合单层和器官型培养物中的同种异体和自体组分。在单层共培养中，NIKSGFP细胞对原代角质形成细胞的生长速率没有影响，并且观察到标记和未标记角质形成细胞之间的细胞-细胞连接形成。在采用真皮和表皮隔室的器官型培养中，使用高达90%的NIKSGFP细胞生成的嵌合复合皮肤替代物表现出正常的组织结构，并具有可归因于原代角质形成细胞的大量区域。组织表达对连续、完全分层的鳞状上皮的结构和功能至关重要的蛋白质，并表现出与天然皮肤相似的屏障功能。此外，嵌合人皮肤替代品在体内小鼠模型中稳定移植，在手术应用后28天，原代角质形成细胞长期保留，但NIKSGFP细胞群丢失。这项研究提供了使用NIKS角质形成细胞作为同种异体细胞来源形成生物工程嵌合皮肤替代组织的概念证明，提供即时的正式伤口覆盖，同时提供自体细胞用于组织再生。
We hypothesize that the pathogen-free human keratinocyte progenitor cell line, NIKS, cultured in a chimeric fashion with patient’s primary keratinocytes would produce a fully-stratified engineered skin substitute tissue and serve to deliver autologous keratinocytes to a cutaneous wound. Chimeric autologous / allogeneic bioengineered skin substitutes offer an innovative regenerative medicine approach for providing wound coverage and restoring cutaneous barrier function while delivering autologous keratinocytes to the wound site. NIKS keratinocytes are an attractive allogeneic cell source for this application. Mixed populations of GFP-labeled NIKS and unlabeled primary keratinocytes were used to model the allogeneic and autologous components in chimeric monolayer and organotypic cultures. In monolayer co-culture, NIKSGFP cells had no effect on the growth rate of primary keratinocytes and cell-cell junction formation between labeled and unlabeled keratinocytes was observed. In organotypic culture employing dermal and epidermal compartments, chimeric composite skin substitutes generated using up to 90% NIKSGFP cells exhibited normal tissue architecture and possessed substantial regions attributable to the primary keratinocytes. Tissues expressed proteins essential for the structure and function of a contiguous, fully-stratified squamous epithelia and exhibited barrier function similar to that of native skin. Furthermore, chimeric human skin substitutes stably engrafted in an in vivo mouse model, with long-term retention of primary keratinocytes but loss of the NIKSGFP cell population by 28 days after surgical application. This study provides proof of concept for the use of NIKS keratinocytes as an allogeneic cell source for the formation of bioengineered chimeric skin substitute tissues, providing immediate formal wound coverage while simultaneously supplying autologous cells for tissue regeneration.