Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS)
Glomerular diseases characterized by injury to post-mitotic epithelial cells called podocytes are a leading cause of chronic kidney disease. Yet, isolating podocytes from the kidney for transcriptomic, proteomic, and metabolomic studies has been a major technical challenge. Protocols utilizing glome...
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Bio-protocol LLC
2025-07-01
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| Series: | Bio-Protocol |
| Online Access: | https://bio-protocol.org/en/bpdetail?id=5364&type=0 |
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| author | Jeffrey Pippin Carol Loretz Diana Eng Oliver Wessely Stuart Shankland |
| author_facet | Jeffrey Pippin Carol Loretz Diana Eng Oliver Wessely Stuart Shankland |
| author_sort | Jeffrey Pippin |
| collection | DOAJ |
| description | Glomerular diseases characterized by injury to post-mitotic epithelial cells called podocytes are a leading cause of chronic kidney disease. Yet, isolating podocytes from the kidney for transcriptomic, proteomic, and metabolomic studies has been a major technical challenge. Protocols utilizing glomerular sieving and laser capture methods are of limited use because they are not podocyte-specific but instead capture all four glomerular cell types. Here, we present a magnetic-activated cell sorting (MACS) method where podocytes are isolated from digested whole kidneys using antibodies specific to extracellular antigens on podocytes. Using microbeaded secondary antibodies binding to the podocyte-specific primary antibodies allows sorting of the podocytes using a magnet. This podocyte-only cell fraction is a unique source of in vivo–derived cells for molecular and cellular experiments. |
| format | Article |
| id | doaj-art-6cf5b85fd12943c7b504de2e95011f7f |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Bio-protocol LLC |
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| spelling | doaj-art-6cf5b85fd12943c7b504de2e95011f7f2025-08-20T03:50:32ZengBio-protocol LLCBio-Protocol2331-83252025-07-01151310.21769/BioProtoc.5364Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS)Jeffrey Pippin0Carol Loretz1Diana Eng2Oliver Wessely3Stuart Shankland4Division of Nephrology, University of Washington, Seattle, WA, USADivision of Nephrology, University of Washington, Seattle, WA, USADivision of Nephrology, University of Washington, Seattle, WA, USALerner Research Institute, Cleveland Clinic Foundation, Cleveland, OH, USADivision of Nephrology, University of Washington, Seattle, WA, USAGlomerular diseases characterized by injury to post-mitotic epithelial cells called podocytes are a leading cause of chronic kidney disease. Yet, isolating podocytes from the kidney for transcriptomic, proteomic, and metabolomic studies has been a major technical challenge. Protocols utilizing glomerular sieving and laser capture methods are of limited use because they are not podocyte-specific but instead capture all four glomerular cell types. Here, we present a magnetic-activated cell sorting (MACS) method where podocytes are isolated from digested whole kidneys using antibodies specific to extracellular antigens on podocytes. Using microbeaded secondary antibodies binding to the podocyte-specific primary antibodies allows sorting of the podocytes using a magnet. This podocyte-only cell fraction is a unique source of in vivo–derived cells for molecular and cellular experiments.https://bio-protocol.org/en/bpdetail?id=5364&type=0 |
| spellingShingle | Jeffrey Pippin Carol Loretz Diana Eng Oliver Wessely Stuart Shankland Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS) Bio-Protocol |
| title | Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS) |
| title_full | Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS) |
| title_fullStr | Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS) |
| title_full_unstemmed | Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS) |
| title_short | Isolation of Podocyte Cell Fractions From Mouse Kidney Using Magnetic Activated Cell Sorting (MACS) |
| title_sort | isolation of podocyte cell fractions from mouse kidney using magnetic activated cell sorting macs |
| url | https://bio-protocol.org/en/bpdetail?id=5364&type=0 |
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