论文标题：Biologically relevant laminin as chemically defined and fully human platform for human epidermal keratinocyte culture
期刊：
作者：Monica Suryana Tjin, Alvin Wen Choong Chua, Aida Moreno-Moral, Li Yen Chong, Po Yin Tang, Nathan Peter Harmston, Zuhua Cai, Enrico Petretto, Bien Keem Tan, Karl Tryggvason
发表时间：2018/10/30
数字识别码： 10.1038/s41467-018-06934-3
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《自然-通讯》论文Biologically relevant laminin as chemically defined and fully human platform for human epidermal keratinocyte culture近日报道了一项最新研发的人体细胞完全培养方法。新方法可产生皮肤移植所需的组织，并有望用于治疗严重烧伤以及其它皮肤问题。
图1：鉴定层粘连蛋白链在人表皮基底膜中的表达。图源：Tjin等
皮肤移植手术近来取得了巨大进步，最多能修复皮肤严重创伤或烧伤患者全身90%的表皮。这种治疗方式需要先从患者身上采集皮肤细胞，将细胞体外培养成一大块组织，再移植到创面上。细胞培养基中通常需要加入小鼠细胞来支持组织生长。但是，这种人体/小鼠细胞混合物会让患者面临发生感染和不良免疫反应的风险。
图2：在层粘连蛋白系统上生长的HEK（人表皮角质形成细胞）的体内功能测定。图源：Tjin 等
杜克-新加坡国立大学明升手机版院的Karl Tryggvason及同事发现，人体中常见的层粘连蛋白的两种特定变体能以类似方式支持皮肤细胞的生长。这一研究结果能让作者在不使用小鼠细胞的情况下，建立无动物源性的人体细胞培养系统，用于皮肤移植。作者随后证实，用这种方法得到的皮肤组织能成功移植到小鼠身上。
摘要：The current expansion of autologous human keratinocytes to resurface severe wound defects still relies on murine feeder layer and calf serum in the cell culture system. Through our characterization efforts of the human skin basement membrane and murine feeder layer 3T3-J2, we identified two biologically relevant recombinant laminins—LN-511 and LN-421- as potential candidates to replace the murine feeder. Herein, we report a completely xeno-free and defined culture system utilizing these laminins which enables robust expansion of adult human skin keratinocytes. We demonstrate that our laminin system is comparable to the 3T3-J2 co-culture system in terms of basal markers’ profile, colony-forming efficiency and the ability to form normal stratified epidermal structure in both in vitro and in vivo models. These results show that the proposed system may not only provide safer keratinocyte use in the clinics, but also facilitate the broader use of other cultured human epithelial cells in regenerative medicine.
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