Du Wenqiang, Hong Sungmin, Scapin Giorgia, Goulard Marie, Shah Dhvanit I
Center for Childhood Cancers and Blood Diseases, Nationwide Children's Hospital, Columbus, Ohio 43205, United States.
Department of Pediatrics, College of Medicine, The Ohio State University, Columbus, Ohio 43210, United States.
ACS Biomater Sci Eng. 2019 Aug 12;5(8):3935-3943. doi: 10.1021/acsbiomaterials.8b01359. Epub 2019 Feb 6.
Directed collective cell migration governs cell orientation during tissue morphogenesis, wound healing, and tumor metastasis. Unfortunately, current methods for initiating collective cell migration, such as scratching, laser ablation, and stencils, either introduce uncontrolled cell-injury, involve multiple fabrication processes, or have utility limited to cells with strong cell-cell junctions. Using three-dimensional (3D) bioprinted gelatin methacryloyl (GelMA) micropatterns on temperature-responsive poly(-isopropylacrylamide) (PNIPAm) coated interfaces, we demonstrate that directed injury-free collective cell migration could occur in parallel and perpendicular directions. After seeding cells, we created cell-free spaces between two 3D bioprinted GelMA micropatterns by lowering the temperature of PNIPAm interfaces to promote the cell detachment. Unlike conventional collective cell migration methods initiated by stencils, we observed well-organized cell migration in parallel and perpendicular to 3D bioprinted micropatterns as a function of the distance between 3D bioprinted micropatterns. We further established the utility of controlled collective cell migration for directed functional myotube formation using 3D bioprinted fingerprintlike micropatterns as well as iris musclelike concentric circular patterns. Our platform is unique for myoblast alignment and myotube formation because it does not require anisotropic guidance cues. Together, our findings establish how to achieve controlled collective cell migration, even at the macroscale, for tissue engineering and regeneration.
定向集体细胞迁移在组织形态发生、伤口愈合和肿瘤转移过程中控制细胞定向。不幸的是，目前启动集体细胞迁移的方法，如划痕、激光消融和模板法，要么会引入不受控制的细胞损伤，涉及多个制造过程，要么其应用仅限于具有强细胞间连接的细胞。通过在温度响应性聚（N-异丙基丙烯酰胺）（PNIPAm）涂层界面上使用三维（3D）生物打印的甲基丙烯酸明胶（GelMA）微图案，我们证明了定向无损伤的集体细胞迁移可以在平行和垂直方向上发生。接种细胞后，我们通过降低PNIPAm界面的温度来促进细胞脱离，从而在两个3D生物打印的GelMA微图案之间创建无细胞空间。与传统的由模板启动的集体细胞迁移方法不同，我们观察到细胞沿着与3D生物打印微图案平行和垂直的方向进行有序迁移，且迁移情况是3D生物打印微图案之间距离的函数。我们进一步利用3D生物打印的指纹状微图案以及虹膜肌样同心圆图案，确立了可控集体细胞迁移在定向功能性肌管形成中的应用。我们的平台对于成肌细胞排列和肌管形成具有独特性，因为它不需要各向异性的引导线索。总之，我们的研究结果确定了如何实现可控的集体细胞迁移，甚至在宏观尺度上，以用于组织工程和再生。
