Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype
Crosstalk between glomerular endothelial cells and glomerular epithelial cells (podocytes) is increasingly becoming apparent as a crucial mechanism to maintain the integrity of the glomerular filtration barrier. However, <i>in vitro</i> studies directly investigating the effect of this c...
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MDPI AG
2023-03-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/13/3/339 |
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| author | Daan C. ‘t Hart Dilemin Yildiz Valentina Palacio-Castañeda Lanhui Li Burcu Gumuscu Roland Brock Wouter P. R. Verdurmen Johan van der Vlag Tom Nijenhuis |
| author_facet | Daan C. ‘t Hart Dilemin Yildiz Valentina Palacio-Castañeda Lanhui Li Burcu Gumuscu Roland Brock Wouter P. R. Verdurmen Johan van der Vlag Tom Nijenhuis |
| author_sort | Daan C. ‘t Hart |
| collection | DOAJ |
| description | Crosstalk between glomerular endothelial cells and glomerular epithelial cells (podocytes) is increasingly becoming apparent as a crucial mechanism to maintain the integrity of the glomerular filtration barrier. However, <i>in vitro</i> studies directly investigating the effect of this crosstalk on the glomerular filtration barrier are scarce because of the lack of suitable experimental models. Therefore, we developed a custom-made glomerulus-on-a-chip model recapitulating the glomerular filtration barrier, in which we investigated the effects of co-culture of glomerular endothelial cells and podocytes on filtration barrier function and the phenotype of these respective cell types. 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. The glycocalyx was visualized by wheat germ agglutinin staining and the barrier integrity of the glomerulus-on-a-chip model was determined by measuring the transport rate of fluorescently labelled dextran from the top to the bottom channel. The effect of crosstalk on the transcriptome of glomerular endothelial cells and podocytes was investigated via RNA-sequencing. Glomerular endothelial cells and podocytes were successfully cultured on opposite sides of the membrane in our glomerulus-on-a-chip model using a polydopamine and collagen A double coating. Barrier integrity of the chip model was significantly improved when glomerular endothelial cells were co-cultured with podocytes compared to monocultures of either glomerular endothelial cells or podocytes. Co-culture enlarged the surface area of podocyte foot processes and increased the thickness of the glycocalyx. RNA-sequencing analysis revealed the regulation of cellular pathways involved in cellular differentiation and cellular adhesion as a result of the interaction between glomerular endothelial cells and podocytes. We present a novel custom-made glomerulus-on-a-chip co-culture model and demonstrated for the first time using a glomerulus-on-a-chip model that co-culture affects the morphology and transcriptional phenotype of glomerular endothelial cells 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. |
| first_indexed | 2024-03-11T06:51:31Z |
| format | Article |
| id | doaj.art-58cfe91930f643149614662cc8ac93c8 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-11T06:51:31Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-58cfe91930f643149614662cc8ac93c82023-11-17T09:53:58ZengMDPI AGBiosensors2079-63742023-03-0113333910.3390/bios13030339Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell PhenotypeDaan C. ‘t Hart0Dilemin Yildiz1Valentina Palacio-Castañeda2Lanhui Li3Burcu Gumuscu4Roland Brock5Wouter P. R. Verdurmen6Johan van der Vlag7Tom Nijenhuis8Department of Nephrology, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsDepartment of Nephrology, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsDepartment of Medical Biosciences, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsBiosensors and Devices Laboratory, Biomedical Engineering Department, Institute for Complex Molecular Systems, Eindhoven Artificial Intelligence Systems Institute, Eindhoven University of Technology, 5600 MB Eindhoven, The NetherlandsBiosensors and Devices Laboratory, Biomedical Engineering Department, Institute for Complex Molecular Systems, Eindhoven Artificial Intelligence Systems Institute, Eindhoven University of Technology, 5600 MB Eindhoven, The NetherlandsDepartment of Medical Biosciences, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsDepartment of Medical Biosciences, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsDepartment of Nephrology, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsDepartment of Nephrology, Research Institute for Medical Innovations, Radboud University Medical Center, 6500 HB Nijmegen, The NetherlandsCrosstalk between glomerular endothelial cells and glomerular epithelial cells (podocytes) is increasingly becoming apparent as a crucial mechanism to maintain the integrity of the glomerular filtration barrier. However, <i>in vitro</i> studies directly investigating the effect of this crosstalk on the glomerular filtration barrier are scarce because of the lack of suitable experimental models. Therefore, we developed a custom-made glomerulus-on-a-chip model recapitulating the glomerular filtration barrier, in which we investigated the effects of co-culture of glomerular endothelial cells and podocytes on filtration barrier function and the phenotype of these respective cell types. 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. The glycocalyx was visualized by wheat germ agglutinin staining and the barrier integrity of the glomerulus-on-a-chip model was determined by measuring the transport rate of fluorescently labelled dextran from the top to the bottom channel. The effect of crosstalk on the transcriptome of glomerular endothelial cells and podocytes was investigated via RNA-sequencing. Glomerular endothelial cells and podocytes were successfully cultured on opposite sides of the membrane in our glomerulus-on-a-chip model using a polydopamine and collagen A double coating. Barrier integrity of the chip model was significantly improved when glomerular endothelial cells were co-cultured with podocytes compared to monocultures of either glomerular endothelial cells or podocytes. Co-culture enlarged the surface area of podocyte foot processes and increased the thickness of the glycocalyx. RNA-sequencing analysis revealed the regulation of cellular pathways involved in cellular differentiation and cellular adhesion as a result of the interaction between glomerular endothelial cells and podocytes. We present a novel custom-made glomerulus-on-a-chip co-culture model and demonstrated for the first time using a glomerulus-on-a-chip model that co-culture affects the morphology and transcriptional phenotype of glomerular endothelial cells 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.https://www.mdpi.com/2079-6374/13/3/339organ-on-a-chipglomerulusglomerulus-on-a-chipco-cultureglomerular filtration barriercrosstalk |
| spellingShingle | Daan C. ‘t Hart Dilemin Yildiz Valentina Palacio-Castañeda Lanhui Li Burcu Gumuscu Roland Brock Wouter P. R. Verdurmen Johan van der Vlag Tom Nijenhuis Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype Biosensors organ-on-a-chip glomerulus glomerulus-on-a-chip co-culture glomerular filtration barrier crosstalk |
| title | Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype |
| title_full | Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype |
| title_fullStr | Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype |
| title_full_unstemmed | Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype |
| title_short | Co-Culture of Glomerular Endothelial Cells and Podocytes in a Custom-Designed Glomerulus-on-a-Chip Model Improves the Filtration Barrier Integrity and Affects the Glomerular Cell Phenotype |
| title_sort | co culture of glomerular endothelial cells and podocytes in a custom designed glomerulus on a chip model improves the filtration barrier integrity and affects the glomerular cell phenotype |
| topic | organ-on-a-chip glomerulus glomerulus-on-a-chip co-culture glomerular filtration barrier crosstalk |
| url | https://www.mdpi.com/2079-6374/13/3/339 |
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