Abstract: PO0492
A 3D Vascularised Tubule Model Improves the Phenotype of Cultured Cells
Session Information
- Bioengineering: Organoids and Organs-on-a-Chip
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Bioengineering
- 300 Bioengineering
Authors
- Carracedo, Miguel, AstraZeneca PLC, Mölndal, Sweden
- Robinson, Sanlin, Harvard University, Wyss Institute, Boston, Massachusetts, United States
- Alaei, Babak, AstraZeneca PLC, Mölndal, Sweden
- Clausen, Maryam, AstraZeneca PLC, Mölndal, Sweden
- Hicks, Ryan, AstraZeneca PLC, Mölndal, Sweden
- Belfield, Graham Paul, AstraZeneca PLC, Mölndal, Sweden
- Bak, Annette, AstraZeneca PLC, Mölndal, Sweden
- Lewis, Jennifer A., Harvard University, Wyss Institute, Boston, Massachusetts, United States
- Laerkegaard Hansen, Pernille B., AstraZeneca PLC, Mölndal, Sweden
- Williams, Julie, AstraZeneca PLC, Mölndal, Sweden
Background
Modelling proximal tubule physiology and pharmacology is essential for mechanistic studies and drug discovery. As a consequence, a plethora of models have been developed. Despite this no comparative study between monocultures, co-cultures and 3D models has been reported. In this study we aimed to understand the differences of proximal tubule epithelial cells (PTECs) and glomerular endothelial cells (HGECs) alone or in co-culture when grown in static non-coated, static matrix-coated and 3D flow matrix-coated conditions.
Methods
We cultured PTECs under physiological flow in a 3D channel embedded within an engineered extracellular matrix (ECM) that is colocalised with an adjacent channel lined with HGECs to mimic a peritubular capillary. After a period of maturation under continuous flow, both cell types were harvested for RNAseq analyses.
Results
Our results revealed that PTECs’ transcriptional profile is highly dependent on the matrix on which these cells are cultured, as well as the application of flow. Endothelial cells however demonstrated greater phenotypic plasticity, being affected by matrix, flow and co-culture. The transcriptional profile of PTECs grown on a non-coated surface presented an enrichment of inflammatory markers such as TNF-a, IL6 and CXCL6, resembling that of diseased tubular biopsies. This inflammatory effect was not seen in PTECs grown on a matrix, and the growth conditions of matrix under flow further resembled the transcriptional profile of healthy tubular biopsies. Unsurprisingly the presence of flow modulated the expression of kidney signature genes including drug/solute transporters. Likewise, HGECs’ transcriptional profile more closely resembled the profile of in vivo glomerular cells when grown on a matrix under flow.
Conclusion
In conclusion, PTECs and HGECs grown under different culture conditions present considerable transcriptional profile changes; being enriched in inflammatory pathways when grown on a non-coated surface, but closely resembling in vivo homeostatic profiles when grown with matrix and/or flow. These findings guide future selection of translatable models investigating renal physiology and pharmacology.
Funding
- Commercial Support –