A microengineered collagen scaffold for generating a polarized crypt-villus architecture of human small intestinal epithelium.
A microengineered collagen scaffold for generating a polarized crypt-villus architecture of human small intestinal epithelium.
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DOI：
10.1016/j.biomaterials.2017.03.005
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发表时间：
2017-06
期刊：
影响因子：
14
通讯作者：
Allbritton NL
中科院分区：
文献类型：
--
作者：
Wang Y;Gunasekara DB;Reed MI;DiSalvo M;Bultman SJ;Sims CE;Magness ST;Allbritton NL
人小肠上皮具有独特的隐窝-绒毛结构和组织极性，其中增殖细胞驻留在隐窝内，而分化细胞定位于绒毛。间接证据表明，分化和迁移的过程是由生化梯度的因素，指定这些细胞室的极性部分驱动;然而，直接证据的梯度驱动的模式，这种在体内的架构已受到体外系统的限制。肠样培养物是一种强大的体外系统;然而，这些球状结构无法复制体内发现的结构和谱系区室化，并且不易于经受生长因子的梯度。在目前的工作中，我们报告了一种具有合适的细胞外基质和刚度的微图案化胶原支架的开发，以产生体外自我更新的人小肠上皮，其复制体内小肠的关键特征：具有适当细胞谱系区室化的隐窝-绒毛结构和开放且可接近的管腔表面。施加到隐窝绒毛轴的化学梯度促进了干/祖细胞区的形成，并支持细胞沿隐窝绒毛轴沿着迁移。这种结合微工程支架、生物物理线索和化学梯度来控制离体肠上皮的新方法可以用作人类小肠上皮的生理相关模拟物，并且广泛适用于模拟依赖于生理功能梯度的其他组织。
The human small intestinal epithelium possesses a distinct crypt-villus architecture and tissue polarity in which proliferative cells reside inside crypts while differentiated cells are localized to the villi. Indirect evidence has shown that the processes of differentiation and migration are driven in part by biochemical gradients of factors that specify the polarity of these cellular compartments; however, direct evidence for gradient-driven patterning of this in vivo architecture has been hampered by limitations of the in vitro systems available. Enteroid cultures are a powerful in vitro system; nevertheless, these spheroidal structures fail to replicate the architecture and lineage compartmentalization found in vivo, and are not easily subjected to gradients of growth factors. In the current work, we report the development of a micropatterned collagen scaffold with suitable extracellular matrix and stiffness to generate an in vitro self-renewing human small intestinal epithelium that replicates key features of the in vivo small intestine: a crypt-villus architecture with appropriate cell-lineage compartmentalization and an open and accessible luminal surface. Chemical gradients applied to the crypt-villus axis promoted the creation of a stem/progenitor-cell zone and supported cell migration along the crypt-villus axis. This new approach combining microengineered scaffolds, biophysical cues and chemical gradients to control the intestinal epithelium ex vivo can serve as a physiologically relevant mimic of the human small intestinal epithelium, and is broadly applicable to model other tissues that rely on gradients for physiological function.