Cultured human keratinocytes on type I collagen membranes to reconstitute the epidermis
Cultured human keratinocytes on type I collagen membranes to reconstitute the epidermis
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DOI：
10.1089/107632700320892
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发表时间：
2000-02-01
影响因子：
--
通讯作者：
Stark, GB
中科院分区：
文献类型：
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作者：
Horch, RE;Debus, M;Stark, GB
开发新技术和改进以克服培养的角质形成细胞移植中的一些缺点是由使用培养的上皮自体移植物中的几个众所周知的缺点所激发的，所述缺点例如长培养期、缺乏粘附性、难以处理、缺乏真皮基质和高成本。最近的两个见解影响了进一步的研究。一方面，已经表明在纤维蛋白胶悬浮液中使用未分化的增殖细胞在上皮重建中是有效的。另一方面，细胞从培养表面的酶促释放是导致培养细胞的锚定结构至少暂时破坏的关键步骤。在本研究中，我们试图将这两个方面联合收割机，试图修改角质形成细胞移植的常见模式。为了避免分散酶溶解培养的细胞，角质形成细胞接种到牛胶原蛋白I型膜上没有饲养层和无血清培养条件下。培养3天后，将培养的人角质形成细胞的亚融合单层作为胶原膜上的倒置移植物（角质形成细胞胶原膜移植物[KCMG]，n = 12）或作为单独的膜移植物（12 = 12）移植到标准裸鼠全层伤口上。KCMG移植后21天表皮完全分化，表皮细胞持续存在。这项研究表明，倒置移植的未分化单层角质形成细胞的非交联牛I型胶原蛋白膜，导致早期重建的多层鳞状上皮，增强伤口愈合相比，对照组。倒置KCMG移植技术能够转移活跃增殖的角质形成细胞，与传统的上皮片移植相比简化了应用。
The development of new techniques and modifications to overcome some of the disadvantages in cultured keratinocyte grafting has been motivated by several well-known drawbacks in the use of cultured epithelial autografts such as long culture periods, lack of adherence, difficulty in handling, lack of dermal substrates, and high costs. Two recent insights have influenced further research. On the one hand, it has been shown that the use of undifferentiated proliferative cells in fibrin glue suspensions is effective in epithelial reconstitution. On the other hand, the enzymatic release of cells from the culture surfaces is a critical step leading to at least temporary destruction of anchoring structures of the cultured cells. In this study, we tried to combine these two aspects in an attempt to modify common modalities of keratinocyte transplantation. To avoid dispase dissolving of the cultured cells, keratinocytes were seeded onto bovine collagen type I membranes without feeder layers and under serum-free culture conditions. Subconfluent monolayers of cultured human keratinocytes were transplanted as an upside-down graft on collagen membranes (keratinocyte collagen membrane grafts [KCMG], n = 12) after 3 days of culture or as membrane grafts alone (12 = 12) onto Standard nude mice full-thickness wounds. Fully differentiated epidermis was found at 21 days after grafting KCMG with persistence of human keratinocytes. This study demonstrates that upside-down grafts of undifferentiated monolayers of keratinocytes on non-cross-linked bovine type I collagen membranes do lead to an early reconstitution of multilayered squamous epithelium with enhanced wound healing compared to the control group. The upside down KCMG grafting technique is able to transfer actively proliferative keratinocytes and simplifies the application compared to conventional epithelial sheet grafting.