RT Journal Article
SR Electronic
T1 Open microfluidic coculture reveals paracrine signaling from human kidney epithelial cells promotes kidney specificity of endothelial cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.02.14.949776
DO 10.1101/2020.02.14.949776
A1 Tianzi Zhang
A1 Daniel Lih
A1 Ryan J. Nagao
A1 Jun Xue
A1 Erwin Berthier
A1 Jonathan Himmelfarb
A1 Ying Zheng
A1 Ashleigh B. Theberge
YR 2020
UL http://biorxiv.org/content/early/2020/03/05/2020.02.14.949776.abstract
AB Endothelial cells (ECs) from different human organs possess organ-specific characteristics that support specific tissue regeneration and organ development. EC specificity are identified by both intrinsic and extrinsic cues, among which, parenchyma and organ-specific microenvironment are critical contributors. These extrinsic cues are, however, largely lost during ex vivo cultures. Outstanding challenges remain to understand and re-establish EC organ-specificity for in vitro studies to recapitulate human organ-specific physiology. Here, we designed an open microfluidic platform to study the role of human kidney tubular epithelial cells in supporting EC specificity. The platform consists of two independent cell culture regions segregated with a half wall; culture media is added to connect the two culture regions at a desired timepoint, and signaling molecules can travel across the half wall (paracrine signaling). Specifically, we report that in the microscale coculture device, primary human kidney proximal tubular epithelial cells (HPTECs) rescued primary human kidney peritubular microvascular EC (HKMEC) monolayer integrity and fenestra formation, and HPTECs upregulated key HKMEC kidney-specific genes (HNF1B, AJAP1, KCNJ16) and endothelial activation genes (VCAM1, MMP7, MMP10) in coculture. Co-culturing with HPTECs also promoted kidney-specific genotype expression in human umbilical vein ECs (HUVECs), and human pluripotent stem cell-derived ECs (hPSC-ECs). In comparison to the culture in HPTEC conditioned media, co-culture of ECs with HPTECs showed increased upregulation of kidney specific genes, suggesting potential bidirectional paracrine signaling. Importantly, our device is compatible with standard pipettes, incubators, and imaging readouts, and could also be easily adapted to study cell signaling between other rare or sensitive cells.