Department of Chemical and Petroleum Engineering, University of Pittsburgh, Pittsburgh, PA, USA.
Lonza, Houston, Texas, PA, USA.
Methods Mol Biol. 2021;2258:73-92. doi: 10.1007/978-1-0716-1174-6_6.
Biophysical cues synergize with biochemical cues to drive differentiation of pluripotent stem cells through specific phenotypic trajectory. Tools to manipulate the cell biophysical environment and identify the influence of specific environment perturbation in the presence of combinatorial inputs will be critical to control the development trajectory. Here we describe the procedure to perturb biophysical environment of pluripotent stem cells while maintaining them in 3D culture configuration. We also discuss a high-throughput platform for combinatorial perturbation of the cell microenvironment, and detail a statistical procedure to extract dominant environmental influences.
生物物理线索与生化线索协同作用，通过特定的表型轨迹驱动多能干细胞的分化。操纵细胞生物物理环境的工具以及识别在组合输入存在下特定环境扰动的影响对于控制发育轨迹将是至关重要的。本文描述了在维持多能干细胞 3D 培养状态的同时，对其生物物理环境进行扰动的方法。我们还讨论了用于细胞微环境组合性扰动的高通量平台，并详细介绍了一种用于提取主要环境影响的统计程序。
