Development of 3D hydrogel culture systems with on-demand cell separation
Development of 3D hydrogel culture systems with on-demand cell separation
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DOI：
10.1002/biot.201200200
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发表时间：
2013-04-01
影响因子：
4.7
通讯作者：
Temenoff, Johnna S.
中科院分区：
文献类型：
--
作者：
Hamilton, Sharon K.;Bloodworth, Nathaniel C.;Temenoff, Johnna S.
最近，人们对细胞群之间的旁分泌信号的作用越来越感兴趣，特别是在更好地理解干细胞如何促进组织修复的背景下。目前大多数3D共培养方法缺乏在培养期后有效分离两个细胞群的能力，这对于同时分析每种细胞类型对另一种细胞类型的相互作用非常重要。在这里，我们详细介绍了3D水凝胶共培养系统的开发，该系统使我们能够培养不同的细胞类型长达7天，随后使用酶敏感胶水分离和分离不同的细胞群。采用酶降解水凝胶粘合剂对聚乙二醇基水凝胶进行层合，制备了可分离的三维共培养层合板。被包裹的细胞群表现出良好的分离，具有良好的界面。此外，在添加适当的酶后，构建物可以按需分离，而细胞活力在整个培养期间保持高水平，即使经过层压分离。该平台为各种基本细胞信号研究提供了巨大的潜力，因为结合了酶敏感的粘附界面，可以按需分离单个细胞群体进行即时分析或进一步培养，以检查共培养效应的持久性和细胞群体的旁分泌信号。参见Danielle R. Bogdanowicz和Helen H. Lu的评论DOI: 10.1002/biot.201300054
Recently there has been an increased interest in the effects of paracrine signaling between groups of cells, particularly in the context of better understanding how stem cells contribute to tissue repair. Most current 3D co-culture methods lack the ability to effectively separate two cell populations after the culture period, which is important for simultaneously analyzing the reciprocal effects of each cell type on the other. Here, we detail the development of a 3D hydrogel co-culture system that allows us to culture different cell types for up to 7 days and subsequently separate and isolate the different cell populations using enzyme-sensitive glues. Separable 3D co-culture laminates were prepared by laminating PEG-based hydrogels with enzyme-degradable hydrogel adhesives. Encapsulated cell populations exhibited good segregation with well-defined interfaces. Furthermore, constructs can be separated on-demand upon addition of the appropriate enzyme, while cell viability remains high throughout the culture period, even after laminate separation. This platform offers great potential for a variety of basic cell signaling studies as the incorporation of an enzyme-sensitive adhesive interface allows the on-demand separation of individual cell populations for immediate analysis or further culture to examine persistence of co-culture effects and paracrine signaling on cell populations. See accompanying commentary by Danielle R. Bogdanowicz and Helen H. Lu DOI: 10.1002/biot.201300054