A Comprehensive Protocol for the Collection, Differentiation, Cryopreservation, and Resuscitation of Primary Murine Bone Marrow Derived Macrophages (BMDM).
| Autor: | Luu AM; Department of Microbiology and Cell Biology, Montana State University, Bozeman, Montana, USA., Shepardson KM; Department of Molecular Cell Biology, University of California Merced, Merced, California, USA., Rynda-Apple A; Department of Microbiology and Cell Biology, Montana State University, Bozeman, Montana, USA. |
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| Jazyk: | angličtina |
| Zdroj: | Immunological investigations [Immunol Invest] 2024 Oct; Vol. 53 (7), pp. 1001-1012. Date of Electronic Publication: 2024 Aug 08. |
| DOI: | 10.1080/08820139.2024.2382805 |
| Abstrakt: | Background: The field of immunology has undoubtedly benefited from the in vitro use of cell lines for immunological studies; however, due to the "immortal" nature of many cell lines, they are not always the best model. Thus, direct collection and culture of primary cells from model organisms is a solution that many researchers utilize. To the best of our knowledge, there is not a singular protocol which encompasses the entire process of bone marrow cell collection through cryopreservation and resuscitation of cells from a murine model. Methods: Bone marrow cells were collected from mice with a C57BL6 genetic background. Cells were differentiated using L929 conditioned media. Cells were assessed using a combination of microscopy, differential staining, immunocytochemistry, and trypan blue. Results: Primary murine BMDMs that underwent cryopreservation followed by resuscitation retained a high degree of viability. Furthermore, these BMDMs retained on overall ability to clear S. aureus . Results: Primary murine BMDMs that underwent cryopreservation followed by resuscitation retained a high degree of viability. Furthermore, these BMDMs retained on overall ability to clear S. aureus . Conclusion: Crypopreserved and resuscitated primary murine BMDMs were viable and retained their pverall S. aureus clearance ability. |
| Databáze: | MEDLINE |
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