Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids
Chronic Kidney Disease (CKD) is a major cause of morbidity and mortality characterized by progressive renal fibrosis, and in extreme cases, renal failure. Human CKD models that replicate the biological complexity of the kidney and CKD are lacking and will be invaluable in identifying drugs to revert...
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MDPI AG
2023-03-01
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| Series: | Organoids |
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| Online Access: | https://www.mdpi.com/2674-1172/2/1/5 |
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| author | Sparshita Nag Ashleigh S. Boyd |
| author_facet | Sparshita Nag Ashleigh S. Boyd |
| author_sort | Sparshita Nag |
| collection | DOAJ |
| description | Chronic Kidney Disease (CKD) is a major cause of morbidity and mortality characterized by progressive renal fibrosis, and in extreme cases, renal failure. Human CKD models that replicate the biological complexity of the kidney and CKD are lacking and will be invaluable in identifying drugs to revert and/or prevent fibrosis. To address this unmet need, we developed 3D renal organoids where human induced pluripotent stem cells (hiPSCs) were differentiated to renal progenitors within a renal extracellular matrix (rECM) gel, based on the premise that an rECM could recreate the renal niche to facilitate hiPSC-derived renal progenitor generation. We used mouse kidneys as a source of rECM and identified that superior detergent-mediated decellularization of mouse kidneys was achieved with a combination of 0.5% <i>w</i>/<i>v</i> Sodium Dodecyl Sulphate and 1% <i>v</i>/<i>v</i> Triton-X and mechanical agitation for 60 h. HiPSCs that underwent specification to become metanephric mesenchyme (MM) were subsequently cultured within the rECM gel and, notably, mesenchymal to epithelial transition (MET) was observed, as judged by expression of nephron markers K-cadherin, Nephrin and WT1. These data demonstrate a role for rECM gel in developing human renal organoids from hiPSCs, which will aid the further development of a human disease model for renal fibrosis. |
| first_indexed | 2024-03-11T06:04:09Z |
| format | Article |
| id | doaj.art-5b308f19fdf0471ea7bdf80862f022af |
| institution | Directory Open Access Journal |
| issn | 2674-1172 |
| language | English |
| last_indexed | 2024-03-11T06:04:09Z |
| publishDate | 2023-03-01 |
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| record_format | Article |
| series | Organoids |
| spelling | doaj.art-5b308f19fdf0471ea7bdf80862f022af2023-11-17T13:08:09ZengMDPI AGOrganoids2674-11722023-03-0121667810.3390/organoids2010005Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal OrganoidsSparshita Nag0Ashleigh S. Boyd1Research Department of Surgical Biotechnology, UCL Division of Surgery & Interventional Science, Royal Free Hospital, Rowland Hill Street, London NW3 2QG, UKResearch Department of Surgical Biotechnology, UCL Division of Surgery & Interventional Science, Royal Free Hospital, Rowland Hill Street, London NW3 2QG, UKChronic Kidney Disease (CKD) is a major cause of morbidity and mortality characterized by progressive renal fibrosis, and in extreme cases, renal failure. Human CKD models that replicate the biological complexity of the kidney and CKD are lacking and will be invaluable in identifying drugs to revert and/or prevent fibrosis. To address this unmet need, we developed 3D renal organoids where human induced pluripotent stem cells (hiPSCs) were differentiated to renal progenitors within a renal extracellular matrix (rECM) gel, based on the premise that an rECM could recreate the renal niche to facilitate hiPSC-derived renal progenitor generation. We used mouse kidneys as a source of rECM and identified that superior detergent-mediated decellularization of mouse kidneys was achieved with a combination of 0.5% <i>w</i>/<i>v</i> Sodium Dodecyl Sulphate and 1% <i>v</i>/<i>v</i> Triton-X and mechanical agitation for 60 h. HiPSCs that underwent specification to become metanephric mesenchyme (MM) were subsequently cultured within the rECM gel and, notably, mesenchymal to epithelial transition (MET) was observed, as judged by expression of nephron markers K-cadherin, Nephrin and WT1. These data demonstrate a role for rECM gel in developing human renal organoids from hiPSCs, which will aid the further development of a human disease model for renal fibrosis.https://www.mdpi.com/2674-1172/2/1/5decellularizationrenal organoidsiPSCsextracellular matrixdirected differentiationmesenchymal to epithelial transition |
| spellingShingle | Sparshita Nag Ashleigh S. Boyd Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids Organoids decellularization renal organoids iPSCs extracellular matrix directed differentiation mesenchymal to epithelial transition |
| title | Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids |
| title_full | Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids |
| title_fullStr | Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids |
| title_full_unstemmed | Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids |
| title_short | Decellularization of Mouse Kidneys to Generate an Extracellular Matrix Gel for Human Induced Pluripotent Stem Cell Derived Renal Organoids |
| title_sort | decellularization of mouse kidneys to generate an extracellular matrix gel for human induced pluripotent stem cell derived renal organoids |
| topic | decellularization renal organoids iPSCs extracellular matrix directed differentiation mesenchymal to epithelial transition |
| url | https://www.mdpi.com/2674-1172/2/1/5 |
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