Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates
Summary: Studying fetal hematopoiesis is challenging as hematopoiesis transitions from the liver to bone marrow. Obtaining human samples is not possible, and small animal models may not provide sufficient biological material. Here, we present a protocol for isolating hematopoietic cells from the non...
| المؤلفون الرئيسيون: | , , , , , , |
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| التنسيق: | مقال |
| اللغة: | English |
| منشور في: |
Elsevier
2024-03-01
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| سلاسل: | STAR Protocols |
| الموضوعات: | |
| الوصول للمادة أونلاين: | http://www.sciencedirect.com/science/article/pii/S2666166724000145 |
| _version_ | 1827356704747028480 |
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| author | Michael J. Nash Evgenia Dobrinskikh Dong Wang Eric M. Pietras Rachel C. Janssen Jacob E. Friedman Stephanie R. Wesolowski |
| author_facet | Michael J. Nash Evgenia Dobrinskikh Dong Wang Eric M. Pietras Rachel C. Janssen Jacob E. Friedman Stephanie R. Wesolowski |
| author_sort | Michael J. Nash |
| collection | DOAJ |
| description | Summary: Studying fetal hematopoiesis is challenging as hematopoiesis transitions from the liver to bone marrow. Obtaining human samples is not possible, and small animal models may not provide sufficient biological material. Here, we present a protocol for isolating hematopoietic cells from the nonhuman primate fetal liver and bone. We describe steps for using cells from the same fetus for fluorescence lifetime imaging microscopy to measure metabolism, assessing cellular function, and flow cytometry for immunophenotyping at the single-cell level.For complete details on the use and execution of this protocol, please refer to Nash et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-08T05:13:48Z |
| format | Article |
| id | doaj.art-1408351a7a1948bd90187a6ca66f9581 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T05:13:48Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-1408351a7a1948bd90187a6ca66f95812024-02-07T04:45:52ZengElsevierSTAR Protocols2666-16672024-03-0151102849Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primatesMichael J. Nash0Evgenia Dobrinskikh1Dong Wang2Eric M. Pietras3Rachel C. Janssen4Jacob E. Friedman5Stephanie R. Wesolowski6Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USADepartment of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USADepartment of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USADepartment of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USAHarold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USAHarold Hamm Diabetes Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA; Corresponding authorDepartment of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA; Corresponding authorSummary: Studying fetal hematopoiesis is challenging as hematopoiesis transitions from the liver to bone marrow. Obtaining human samples is not possible, and small animal models may not provide sufficient biological material. Here, we present a protocol for isolating hematopoietic cells from the nonhuman primate fetal liver and bone. We describe steps for using cells from the same fetus for fluorescence lifetime imaging microscopy to measure metabolism, assessing cellular function, and flow cytometry for immunophenotyping at the single-cell level.For complete details on the use and execution of this protocol, please refer to Nash et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166724000145Cell-based AssaysImmunologyMetabolismModel Organisms |
| spellingShingle | Michael J. Nash Evgenia Dobrinskikh Dong Wang Eric M. Pietras Rachel C. Janssen Jacob E. Friedman Stephanie R. Wesolowski Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates STAR Protocols Cell-based Assays Immunology Metabolism Model Organisms |
| title | Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates |
| title_full | Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates |
| title_fullStr | Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates |
| title_full_unstemmed | Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates |
| title_short | Isolating mononuclear cells from fetal bone and liver for metabolic, functional, and immunophenotypic analyses in nonhuman primates |
| title_sort | isolating mononuclear cells from fetal bone and liver for metabolic functional and immunophenotypic analyses in nonhuman primates |
| topic | Cell-based Assays Immunology Metabolism Model Organisms |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724000145 |
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