3D vascularized proximal tubule model improves cell phenotype

Cultured proximal tubule epithelial cells (PTECs) under physiological flow in a 3D channel embedded within an engineered extracellular matrix (ECM) that is colocalized with an adjacent channel lined with human glomerular endothelial cells (HGECs) to mimic a peritubular capillary. After a period of maturation under continuous flow, both cell types were harvested for RNAseq analyses. Our results revealed that PTECs’ transcriptional profile is highly dependent on both the matrix on which these cells are cultured and the flow conditions. By contrast, endothelial cells exhibit greater phenotypic plasticity and are affected by matrix, flow, and co-culture conditions. PTECs and HGECs were grown alone or in co-culture in three different condition: plastic, embedded within an engineered extracellular matrix, or embedded within an engineered extracellular matrix under flow. RNA was extracted and RNA-seq performed.

本数据集对应的实验如下：将培养的近端肾小管上皮细胞（proximal tubule epithelial cells，PTECs）置于嵌入工程化细胞外基质（extracellular matrix，ECM）的三维通道内，于生理流体条件下培养，并使其与由人肾小球内皮细胞（human glomerular endothelial cells，HGECs）衬里的相邻通道毗邻，以此模拟肾小管周毛细血管。经过持续流体环境下的成熟培养周期后，收集两种细胞以开展RNA测序（RNA-seq）分析。研究结果显示，PTECs的转录组谱高度依赖于细胞培养所用基质类型与流体培养条件。与之相对，内皮细胞展现出更强的表型可塑性，其表型受基质、流体条件以及共培养环境的共同调控。本实验设置了三类培养条件：分别以单独培养或共培养的方式，将细胞培养于塑料培养表面、嵌入工程化细胞外基质中，以及在流体条件下嵌入工程化细胞外基质中。随后提取细胞总RNA并完成RNA测序。