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Abstract: SA-PO011

Development of a Glomerulus-on-a-Chip Model: An Innovative Model to Study Paracrine Pathways to Unravel Glomerular Disorders

Session Information

Category: Bioengineering

  • 400 Bioengineering

Authors

  • Yildiz, Dilemin, Radboudumc, Nijmegen, Gelderland, Netherlands
  • 't Hart, Daan C., Radboudumc, Nijmegen, Gelderland, Netherlands
  • Di Giovanni, Gianluca Vincenzo, Radboudumc, Nijmegen, Gelderland, Netherlands
  • Gumuscu, Burcu, Technische Universiteit Eindhoven, Eindhoven, Noord-Brabant, Netherlands
  • Van der vlag, Johan, Radboudumc, Nijmegen, Gelderland, Netherlands
  • Nijenhuis, Tom, Radboudumc, Nijmegen, Gelderland, Netherlands
Background

Crosstalk between glomerular endothelial cells (GEnC) and glomerular epithelial cells (podocytes) is increasingly becoming apparent as a crucial mechanism to maintain the glomerular filtration barrier (GFB) integrity. However, in vitro studies directly investigating the effect of this crosstalk are scarce because of the lack of suitable models. Therefore, we developed a custom-made glomerulus-on-a-chip model recapitulating the GFB, in which we investigated the effects of co-culture of GEnC and podocytes on barrier function and cellular phenotype.

Methods

The custom-made glomerulus-on-a-chip model was designed using soft lithography. The chip consisted of two parallel microfluidic channels separated by a semi-permeable polycarbonate membrane. iPSC derived podocytes and conditionally immortalized GEnC and podocytes were differentiated in the chip. Shear stress induced by laminar flow was applied on endothelial cells. Cellular morphology and glycocalyx was visualized by fluorescent staining. The barrier integrity of the model was determined by measuring the transport rate of labelled dextran from top to bottom channel. The effect of crosstalk on the transcriptome of GEnC and podocytes was investigated via RNA-sequencing.

Results

GEnC and podocytes were successfully cultured on opposite sides of the membrane in our glomerulus-on-a-chip. Barrier integrity of the chip was significantly improved when GEnC were co-cultured with podocytes compared to monocultures. Co-culture enlarged podocyte foot process surface area and increased glycocalyx thickness. RNA-sequencing analysis revealed the regulation of cellular pathways involved in cellular differentiation and cellular adhesion as a result of the interaction between GEnC and podocytes.

Conclusion

We present a novel custom-made glomerulus-on-a-chip and demonstrated that co-culture in the device affects the morphology and transcriptional phenotype of GEnC and podocytes. Moreover, we showed that co-culture improves barrier function as a relevant functional readout for clinical translation. This model can be used in future studies to investigate specific glomerular paracrine pathways and unravel the role of glomerular crosstalk in glomerular (patho) physiology.