O'Grady Brian J, Balikov Daniel A, Lippmann Ethan S, Bellan Leon M
Department of Mechanical Engineering, Vanderbilt University, Nashville, Tennessee.
Interdisciplinary Materials Science Program, Vanderbilt University, Nashville, Tennessee.
Curr Protoc Stem Cell Biol. 2019 Dec;51(1):e97. doi: 10.1002/cpsc.97.
Morphogens are biological molecules that alter cellular identity and behavior across both space and time. During embryonic development, morphogen spatial localization can be confined to small volumes in a single tissue or permeate throughout an entire organism, and the temporal effects of morphogens can range from fractions of a second to several days. In most cases, morphogens are presented as a gradient to adjacent cells within tissues to pattern cell fate. As such, to appropriately model development and build representative multicellular architectures in vitro, it is vital to recapitulate these gradients during stem cell differentiation. However, the ability to control morphogen presentation within in vitro systems remains challenging. Here, we describe an innovative platform using channels patterned within thick, three-dimensional hydrogels that deliver multiple morphogens to embedded cells, thereby demonstrating exquisite control over both spatial and temporal variations in morphogen presentation. This generalizable approach should have broad utility for researchers interested in patterning in vitro tissue structures. © 2019 by John Wiley & Sons, Inc.
形态发生素是一类生物分子,能够在空间和时间维度上改变细胞特性与行为。在胚胎发育过程中,形态发生素的空间定位既可以局限于单个组织中的小区域,也可以遍布整个生物体,其时间效应则可以从几分之一秒到数天不等。在大多数情况下,形态发生素以梯度形式呈现给组织内的相邻细胞,从而决定细胞命运。因此,为了在体外恰当地模拟发育过程并构建具有代表性的多细胞结构,在干细胞分化过程中重现这些梯度至关重要。然而,在体外系统中控制形态发生素的呈现能力仍然具有挑战性。在此,我们描述了一个创新平台,该平台利用在厚的三维水凝胶中形成图案的通道,将多种形态发生素传递给包埋其中的细胞,从而展示了对形态发生素呈现的空间和时间变化的精确控制。这种通用方法对于对体外组织结构图案化感兴趣的研究人员应该具有广泛的用途。© 2019 约翰·威利父子公司版权所有