Regenerative Biology, Morgridge Institute for Research, Madison, WI 53715, USA.
Department of Biomedical Engineering, University of Wisconsin-Madison, Wisconsin Institute for Medical Research, Room 5405, Madison, WI 53706, USA.
Stem Cell Reports. 2017 Apr 11;8(4):907-918. doi: 10.1016/j.stemcr.2017.02.014. Epub 2017 Mar 23.
A defined protocol for efficiently deriving endothelial cells from human pluripotent stem cells was established and vascular morphogenesis was used as a model system to understand how synthetic hydrogels influence global biological function compared with common 2D and 3D culture platforms. RNA sequencing demonstrated that gene expression profiles were similar for endothelial cells and pericytes cocultured in polyethylene glycol (PEG) hydrogels or Matrigel, while monoculture comparisons identified distinct vascular signatures for each cell type. Endothelial cells cultured on tissue-culture polystyrene adopted a proliferative phenotype compared with cells cultured on or encapsulated in PEG hydrogels. The proliferative phenotype correlated to increased FAK-ERK activity, and knockdown or inhibition of ERK signaling reduced proliferation and expression for cell-cycle genes while increasing expression for "3D-like" vasculature development genes. Our results provide insight into the influence of 2D and 3D culture formats on global biological processes that regulate cell function.
建立了一种从人多能干细胞中高效获得内皮细胞的定义明确的方案，并将血管形态发生用作模型系统，以了解与常见的 2D 和 3D 培养平台相比，合成水凝胶如何影响全局生物学功能。RNA 测序表明，共培养在聚乙二醇（PEG）水凝胶或 Matrigel 中的内皮细胞和周细胞的基因表达谱相似，而单核培养比较则为每种细胞类型鉴定了独特的血管特征。与培养在 PEG 水凝胶上或包封在 PEG 水凝胶中的细胞相比，在组织培养聚苯乙烯上培养的内皮细胞表现出增殖表型。增殖表型与 FAK-ERK 活性增加相关，ERK 信号转导的敲低或抑制减少了细胞周期基因的增殖和表达，同时增加了“3D 样”血管发育基因的表达。我们的结果提供了对调节细胞功能的 2D 和 3D 培养形式对全局生物学过程的影响的深入了解。
