Relaxation of Extracellular Matrix Forces Directs Crypt Formation and Architecture in Intestinal Organoids
Relaxation of Extracellular Matrix Forces Directs Crypt Formation and Architecture in Intestinal Organoids
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DOI：
10.1002/adhm.201901214
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发表时间：
2020-01-20
影响因子：
10
通讯作者：
Anseth, Kristi S.
中科院分区：
文献类型：
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作者：
Hushka, Ella A.;Yavitt, F. Max;Anseth, Kristi S.
肠类器官方案依赖于使用细胞外支架，通常是基质胶，并且在从生长切换到分化促进培养基时，发生对称性破坏事件。在此阶段，第一个芽状结构类似于隐窝从中央体伸出，分化开始。虽然类器官提供了无与伦比的结构和功能复杂性，但这种复杂性也是均匀的隐窝-绒毛结构的高度可变性和缺乏再现性的原因。如果在类器官中功能遵循形式，则这种结构变异性对翻译应用具有潜在的限制（例如，药物筛选）。因此，人们对开发合成生物材料以指导类器官生长和分化感兴趣。有人假设合成支架软化是隐窝发育所必需的，这些机械要求提出了一个问题，什么样的压力和随后的松弛是类器官成熟所必需的？为此，烯丙基硫化物水凝胶被用作合成的细胞外基质模拟物，但具有在时间上调节材料的体积模量的光可裂解键。通过改变基质软化的程度，它表明，隐窝的形成，大小，和每菌落数是基质软化的函数。对隐窝结构的机械依赖性的理解对于指导用于临床应用的同质的、可再现的类器官是必要的。
Intestinal organoid protocols rely on the use of extracellular scaffolds, typically Matrigel, and upon switching from growth to differentiation promoting media, a symmetry breaking event takes place. During this stage, the first bud like structures analogous to crypts protrude from the central body and differentiation ensues. While organoids provide unparalleled architectural and functional complexity, this sophistication is also responsible for the high variability and lack of reproducibility of uniform crypt-villus structures. If function follows form in organoids, such structural variability carries potential limitations for translational applications (e.g., drug screening). Consequently, there is interest in developing synthetic biomaterials to direct organoid growth and differentiation. It has been hypothesized that synthetic scaffold softening is necessary for crypt development, and these mechanical requirements raise the question, what compressive forces and subsequent relaxation are necessary for organoid maturation? To that end, allyl sulfide hydrogels are employed as a synthetic extracellular matrix mimic, but with photocleavable bonds that temporally regulate the material's bulk modulus. By varying the extent of matrix softening, it is demonstrated that crypt formation, size, and number per colony are functions of matrix softening. An understanding of the mechanical dependence of crypt architecture is necessary to instruct homogenous, reproducible organoids for clinical applications.