Rapid tissue engineering of biomimetic human corneal limbal crypts with 3D niche architecture
Rapid tissue engineering of biomimetic human corneal limbal crypts with 3D niche architecture
复制标题
DOI：
10.1016/j.biomaterials.2013.08.002
复制
发表时间：
2013-11-01
期刊：
影响因子：
14
通讯作者：
Daniels, Julie T.
中科院分区：
文献类型：
--
作者：
Levis, Hannah J.;Massie, Isobel;Daniels, Julie T.
角膜缘上皮干细胞负责人角膜上皮的维持，并且这些细胞驻留在专门的干细胞龛中。它们位于角膜缘隐窝的底部，在物理保护的微环境中，靠近各种相邻的小生境细胞。设计和重建各种干细胞小生境的元素使研究人员能够简化这些复杂微环境的各个方面，以便进一步进行体外研究。我们已经开发了一种方法，使用简单的一步法在胶原蛋白构建体中快速且可重复地创建生物工程角膜缘隐窝（BLC）。使用亲水性多孔吸收剂（HPA）从胶原蛋白水凝胶中去除液体，所述亲水性多孔吸收剂在基底上具有定制的模制微脊。薄胶原构建物表面上的所得形貌类似于人利姆布斯的基质隐窝的尺寸。接种在构建体表面上的人角膜缘上皮细胞填充这些BLC并形成许多层，其中高比例的细胞衬在表达推定的干细胞标记物p63 α的隐窝中。HPAs采用灵活的成型工艺生产，可根据最终用户的要求进行调整。使用该过程创建定义的地形特征可以适用于需要不同的3维生态位架构的许多组织工程应用。(C)2013爱思唯尔有限公司保留所有权利。
Limbal epithelial stem cells are responsible for the maintenance of the human corneal epithelium and these cells reside in a specialised stem cell niche. They are located at the base of limbal crypts, in a physically protected microenvironment in close proximity to a variety of neighbouring niche cells. Design and recreation of elements of various stem cell niches have allowed researchers to simplify aspects of these complex microenvironments for further study in vitro. We have developed a method to rapidly and reproducibly create bioengineered limbal crypts (BLCs) in a collagen construct using a simple one-step method. Liquid is removed from collagen hydrogels using hydrophilic porous absorbers (HPAs) that have custom moulded micro-ridges on the base. The resulting topography on the surface of the thin collagen constructs resembles the dimensions of the stromal crypts of the human limbus. Human limbal epithelial cells seeded onto the surface of the constructs populate these BLCs and form numerous layers with a high proportion of the cells lining the crypts expressing putative stem cell marker, p63 alpha. The HPAs are produced using a moulding process that is flexible and can be adapted depending on the requirements of the end user. Creation of defined topographical features using this process could be applicable to numerous tissue-engineering applications where varied 3-dimensional niche architectures are required. (C) 2013 Elsevier Ltd. All rights reserved.