Cargando…
Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres
BACKGROUND: Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite the implications for regenerative medicine, their cultivation and expansion remain challenging. Primary explant cell c...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10617125/ https://www.ncbi.nlm.nih.gov/pubmed/37904232 http://dx.doi.org/10.1186/s13287-023-03529-8 |
_version_ | 1785129538101968896 |
---|---|
author | Ocampo-Godinez, Juan M. Gonzalez-Quiroz, Jose L. Cote-Palafox, Hector George, Elizabeth Vergara-Lope Nuñez, Jael A. Villagomez-Olea, Guillermo Vazquez-Vazquez, Febe C. Lopez-Villegas, Edgar O. Leon-Avila, Gloria Dominguez-Lopez, Maria L. Alvarez-Perez, Marco A. |
author_facet | Ocampo-Godinez, Juan M. Gonzalez-Quiroz, Jose L. Cote-Palafox, Hector George, Elizabeth Vergara-Lope Nuñez, Jael A. Villagomez-Olea, Guillermo Vazquez-Vazquez, Febe C. Lopez-Villegas, Edgar O. Leon-Avila, Gloria Dominguez-Lopez, Maria L. Alvarez-Perez, Marco A. |
author_sort | Ocampo-Godinez, Juan M. |
collection | PubMed |
description | BACKGROUND: Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite the implications for regenerative medicine, their cultivation and expansion remain challenging. Primary explant cell culture is a technique that allows the seeding and expansion of difficult-to-culture cells. Here, we report a reliable and simple culture system to obtain functional TECs and thymic interstitial cells (TICs). METHODS: To establish primary thymic explants, we harvested 1 mm cleaned fragments of thymus from 5-week-old C57/BL6 mice. Tissue fragments of a complete thymic lobe were placed in the center of a Petri dish with 1 mL of DMEM/F-12 medium supplemented with 20% fetal bovine serum (FBS) and 1% penicillin‒streptomycin. To compare, thymic explants were also cultivated by using serum-free DMEM/F-12 medium supplemented with 10% KnockOut™. RESULTS: We obtained high numbers of functional clonogenic TECs and TICs from primary thymic explants cultivated with DMEM/F-12 with 20% FBS. These cells exhibited a highly proliferative and migration profile and were able to constitute thymospheres. Furthermore, all the subtypes of medullary TECs were identified in this system. They express functional markers to shape T-cell and type 2 innate lymphoid cells repertoires, such as Aire, IL25, CCL21 and CD80. Finally, we also found that ≥ 70% of lineage negative TICs expressed high amounts of Aire and IL25. CONCLUSION: Thymic explants are an efficient method to obtain functional clonogenic TECs, all mTEC subsets and different TICs Aire(+)IL25(+) with high regenerative capacity. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-023-03529-8. |
format | Online Article Text |
id | pubmed-10617125 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-106171252023-11-01 Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres Ocampo-Godinez, Juan M. Gonzalez-Quiroz, Jose L. Cote-Palafox, Hector George, Elizabeth Vergara-Lope Nuñez, Jael A. Villagomez-Olea, Guillermo Vazquez-Vazquez, Febe C. Lopez-Villegas, Edgar O. Leon-Avila, Gloria Dominguez-Lopez, Maria L. Alvarez-Perez, Marco A. Stem Cell Res Ther Short Report BACKGROUND: Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite the implications for regenerative medicine, their cultivation and expansion remain challenging. Primary explant cell culture is a technique that allows the seeding and expansion of difficult-to-culture cells. Here, we report a reliable and simple culture system to obtain functional TECs and thymic interstitial cells (TICs). METHODS: To establish primary thymic explants, we harvested 1 mm cleaned fragments of thymus from 5-week-old C57/BL6 mice. Tissue fragments of a complete thymic lobe were placed in the center of a Petri dish with 1 mL of DMEM/F-12 medium supplemented with 20% fetal bovine serum (FBS) and 1% penicillin‒streptomycin. To compare, thymic explants were also cultivated by using serum-free DMEM/F-12 medium supplemented with 10% KnockOut™. RESULTS: We obtained high numbers of functional clonogenic TECs and TICs from primary thymic explants cultivated with DMEM/F-12 with 20% FBS. These cells exhibited a highly proliferative and migration profile and were able to constitute thymospheres. Furthermore, all the subtypes of medullary TECs were identified in this system. They express functional markers to shape T-cell and type 2 innate lymphoid cells repertoires, such as Aire, IL25, CCL21 and CD80. Finally, we also found that ≥ 70% of lineage negative TICs expressed high amounts of Aire and IL25. CONCLUSION: Thymic explants are an efficient method to obtain functional clonogenic TECs, all mTEC subsets and different TICs Aire(+)IL25(+) with high regenerative capacity. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-023-03529-8. BioMed Central 2023-10-31 /pmc/articles/PMC10617125/ /pubmed/37904232 http://dx.doi.org/10.1186/s13287-023-03529-8 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Short Report Ocampo-Godinez, Juan M. Gonzalez-Quiroz, Jose L. Cote-Palafox, Hector George, Elizabeth Vergara-Lope Nuñez, Jael A. Villagomez-Olea, Guillermo Vazquez-Vazquez, Febe C. Lopez-Villegas, Edgar O. Leon-Avila, Gloria Dominguez-Lopez, Maria L. Alvarez-Perez, Marco A. Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
title | Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
title_full | Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
title_fullStr | Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
title_full_unstemmed | Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
title_short | Primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
title_sort | primary explants of the postnatal thymus allow the expansion of clonogenic thymic epithelial cells that constitute thymospheres |
topic | Short Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10617125/ https://www.ncbi.nlm.nih.gov/pubmed/37904232 http://dx.doi.org/10.1186/s13287-023-03529-8 |
work_keys_str_mv | AT ocampogodinezjuanm primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT gonzalezquirozjosel primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT cotepalafoxhector primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT georgeelizabeth primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT vergaralopenunezjaela primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT villagomezoleaguillermo primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT vazquezvazquezfebec primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT lopezvillegasedgaro primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT leonavilagloria primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT dominguezlopezmarial primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres AT alvarezperezmarcoa primaryexplantsofthepostnatalthymusallowtheexpansionofclonogenicthymicepithelialcellsthatconstitutethymospheres |