Leight Jennifer L, Liu Wendy F, Chaturvedi Ritika R, Chen Sophia, Yang Michael T, Raghavan Srivatsan, Chen Christopher S
Department of Bioengineering, University of Pennsylvania, 510 Skirkanich Hall, 210 South 33rd Street, Philadelphia, PA 19104, USA.
Cell Mol Bioeng. 2012 Sep 1;5(3):299-306. doi: 10.1007/s12195-012-0236-9.
A novel method to control three-dimensional cell cluster size and geometry using two-dimensional patterning techniques is described. Cells were first cultured on two-dimensional micropatterned collagen using conventional soft lithography techniques. Collagenase was used to degrade the micropatterned collagen and release cells from the micropatterns, forming clusters of cells which were then resuspended in a three-dimensional collagen matrix. This method facilitated the formation of uniformly sized clusters within a single sample. By systematically varying the geometry of the two-dimensional micropatterned islands, final cluster size and cell number in three dimensions could be controlled. Using this technique, we showed that proliferation of cells within collagen gels depended on the size of clusters, suggesting an important role for multicellular structure on biological function. Furthermore, by utilizing more complex two-dimensional patterns, non-spherical structures could be produced. This technique demonstrates a simple way to exploit two-dimensional micro-patterning in order to create complex and structured multicellular clusters in a three-dimensional environment.
描述了一种使用二维图案化技术控制三维细胞簇大小和几何形状的新方法。首先使用传统的软光刻技术在二维微图案化胶原蛋白上培养细胞。使用胶原酶降解微图案化胶原蛋白并将细胞从微图案中释放出来，形成细胞簇，然后将其重悬于三维胶原蛋白基质中。该方法有助于在单个样本中形成大小均匀的簇。通过系统地改变二维微图案化岛的几何形状，可以控制三维中的最终簇大小和细胞数量。使用该技术，我们表明胶原蛋白凝胶内细胞的增殖取决于簇的大小，这表明多细胞结构对生物学功能具有重要作用。此外，通过利用更复杂的二维图案，可以产生非球形结构。该技术展示了一种利用二维微图案化来在三维环境中创建复杂且结构化的多细胞簇的简单方法。
