Phagocytosis model of calcium oxalate monohydrate crystals generated using human induced pluripotent stem cell-derived macrophages
© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature..
Macrophages play a role in nephrolithiasis, offering the possibility of developing macrophage-mediated preventive therapies. To establish a system for screening drugs that could prevent the formation of kidney stones, we aimed to develop a model using human induced pluripotent stem cell (iPSC)-derived macrophages to study phagocytosis of calcium oxalate monohydrate (COM) crystals. Human iPSCs (201B7) were cultured. CD14+ monocytes were recovered using a stepwise process that involved the use of growth factors and cytokines. These cells were then allowed to differentiate into M1 and M2 macrophages. The macrophages were co-cultured with COM crystals and used in the phagocytosis experiments. Live cell imaging and polarized light observation via super-resolution microscopy were used to visualize phagocytosis. Localization of phagocytosed COM crystals was observed using transmission electron microscopy. Intracellular fluorescence intensity was measured using imaging cytometry to quantify phagocytosis. Human iPSCs successfully differentiated into M1 and M2 macrophages. M1 macrophages adhered to the culture plate and moved COM crystals from the periphery to cell center over time, whereas M2 macrophages did not adhere to the culture plate and actively phagocytosed the surrounding COM crystals. Fluorescence assessment over a 24-h period showed that M2 macrophages exhibited higher intracellular fluorescence intensity (5.65-times higher than that of M1 macrophages at 4.5 h) and maintained this advantage for 18 h. This study revealed that human iPSC-derived macrophages have the ability to phagocytose COM crystals, presenting a new approach for studying urinary stone formation and highlighting the potential of iPSC-derived macrophages as a tool to screen nephrolithiasis-related drugs.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:52 |
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| Enthalten in: |
Urolithiasis - 52(2024), 1 vom: 30. März, Seite 51 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Okada, Tomoki [VerfasserIn] |
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| Links: |
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| Themen: |
2612HC57YE |
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| Anmerkungen: |
Date Completed 01.04.2024 Date Revised 01.04.2024 published: Electronic Citation Status MEDLINE |
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| doi: |
10.1007/s00240-024-01553-8 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM370436903 |
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| 520 | |a Macrophages play a role in nephrolithiasis, offering the possibility of developing macrophage-mediated preventive therapies. To establish a system for screening drugs that could prevent the formation of kidney stones, we aimed to develop a model using human induced pluripotent stem cell (iPSC)-derived macrophages to study phagocytosis of calcium oxalate monohydrate (COM) crystals. Human iPSCs (201B7) were cultured. CD14+ monocytes were recovered using a stepwise process that involved the use of growth factors and cytokines. These cells were then allowed to differentiate into M1 and M2 macrophages. The macrophages were co-cultured with COM crystals and used in the phagocytosis experiments. Live cell imaging and polarized light observation via super-resolution microscopy were used to visualize phagocytosis. Localization of phagocytosed COM crystals was observed using transmission electron microscopy. Intracellular fluorescence intensity was measured using imaging cytometry to quantify phagocytosis. Human iPSCs successfully differentiated into M1 and M2 macrophages. M1 macrophages adhered to the culture plate and moved COM crystals from the periphery to cell center over time, whereas M2 macrophages did not adhere to the culture plate and actively phagocytosed the surrounding COM crystals. Fluorescence assessment over a 24-h period showed that M2 macrophages exhibited higher intracellular fluorescence intensity (5.65-times higher than that of M1 macrophages at 4.5 h) and maintained this advantage for 18 h. This study revealed that human iPSC-derived macrophages have the ability to phagocytose COM crystals, presenting a new approach for studying urinary stone formation and highlighting the potential of iPSC-derived macrophages as a tool to screen nephrolithiasis-related drugs | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Calcium oxalate monohydrate | |
| 650 | 4 | |a Cytokine | |
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| 700 | 1 | |a Onozato, Daichi |e verfasserin |4 aut | |
| 700 | 1 | |a Kato, Taiki |e verfasserin |4 aut | |
| 700 | 1 | |a Takase, Hiroshi |e verfasserin |4 aut | |
| 700 | 1 | |a Ohshima, Shigeru |e verfasserin |4 aut | |
| 700 | 1 | |a Sugino, Teruaki |e verfasserin |4 aut | |
| 700 | 1 | |a Unno, Rei |e verfasserin |4 aut | |
| 700 | 1 | |a Taguchi, Kazumi |e verfasserin |4 aut | |
| 700 | 1 | |a Hamamoto, Shuzo |e verfasserin |4 aut | |
| 700 | 1 | |a Ando, Ryosuke |e verfasserin |4 aut | |
| 700 | 1 | |a Shimada, Issei S |e verfasserin |4 aut | |
| 700 | 1 | |a Hashita, Tadahiro |e verfasserin |4 aut | |
| 700 | 1 | |a Iwao, Takahiro |e verfasserin |4 aut | |
| 700 | 1 | |a Matsunaga, Tamihide |e verfasserin |4 aut | |
| 700 | 1 | |a Yasui, Takahiro |e verfasserin |4 aut | |
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