Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system
Summary: This protocol describes an ex vivo cell culture system for simultaneously generating a mixture of CD4+ T helper lineages, including T helper 17 (Th17), RORγt+ Treg, and conventional Treg (cTreg), in proportions representative of those found in mucosal tissues in vivo. When combined with a c...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722004233 |
| _version_ | 1818198258002952192 |
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| author | Shengyun Ma Juan E. Hernandez Wendy Jia Men Huang |
| author_facet | Shengyun Ma Juan E. Hernandez Wendy Jia Men Huang |
| author_sort | Shengyun Ma |
| collection | DOAJ |
| description | Summary: This protocol describes an ex vivo cell culture system for simultaneously generating a mixture of CD4+ T helper lineages, including T helper 17 (Th17), RORγt+ Treg, and conventional Treg (cTreg), in proportions representative of those found in mucosal tissues in vivo. When combined with a co-culture approach, this system allows a more rapid assessment of a candidate molecule’s T cell-intrinsic and -extrinsic functions over the traditional bone marrow chimera and co-transfer approaches.For complete details on the use and execution of this protocol, please refer to Ma et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-12-12T02:03:00Z |
| format | Article |
| id | doaj.art-a51f10e38c994fc6a2939836997e1072 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-12T02:03:00Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-a51f10e38c994fc6a2939836997e10722022-12-22T00:42:08ZengElsevierSTAR Protocols2666-16672022-09-0133101543Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture systemShengyun Ma0Juan E. Hernandez1Wendy Jia Men Huang2Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093, USA; Corresponding authorDepartment of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093, USADepartment of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA 92093, USA; Corresponding authorSummary: This protocol describes an ex vivo cell culture system for simultaneously generating a mixture of CD4+ T helper lineages, including T helper 17 (Th17), RORγt+ Treg, and conventional Treg (cTreg), in proportions representative of those found in mucosal tissues in vivo. When combined with a co-culture approach, this system allows a more rapid assessment of a candidate molecule’s T cell-intrinsic and -extrinsic functions over the traditional bone marrow chimera and co-transfer approaches.For complete details on the use and execution of this protocol, please refer to Ma et al. (2022). : 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/S2666166722004233Cell BiologyCell cultureFlow Cytometry/Mass CytometryImmunology |
| spellingShingle | Shengyun Ma Juan E. Hernandez Wendy Jia Men Huang Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system STAR Protocols Cell Biology Cell culture Flow Cytometry/Mass Cytometry Immunology |
| title | Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system |
| title_full | Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system |
| title_fullStr | Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system |
| title_full_unstemmed | Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system |
| title_short | Protocol to assess cell-intrinsic regulatory mechanisms using an ex vivo murine T cell polarization and co-culture system |
| title_sort | protocol to assess cell intrinsic regulatory mechanisms using an ex vivo murine t cell polarization and co culture system |
| topic | Cell Biology Cell culture Flow Cytometry/Mass Cytometry Immunology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722004233 |
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