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In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells

We report a serum-free, 3D murine artificial thymic organoid (M-ATO) system that mimics normal murine thymopoiesis with the production of all T cell stages, from early thymic progenitors to functional single-positive (CD8SP and CD4SP) TCRαβ and TCRγδ cells. RNA sequencing aligns M-ATO-derived popula...

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Detalles Bibliográficos
Autores principales: Montel-Hagen, Amélie, Sun, Victoria, Casero, David, Tsai, Steven, Zampieri, Alexandre, Jackson, Nicholas, Li, Suwen, Lopez, Shawn, Zhu, Yuhua, Chick, Brent, He, Chongbin, de Barros, Stéphanie C., Seet, Christopher S., Crooks, Gay M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7727762/
https://www.ncbi.nlm.nih.gov/pubmed/33113379
http://dx.doi.org/10.1016/j.celrep.2020.108320
Descripción
Sumario:We report a serum-free, 3D murine artificial thymic organoid (M-ATO) system that mimics normal murine thymopoiesis with the production of all T cell stages, from early thymic progenitors to functional single-positive (CD8SP and CD4SP) TCRαβ and TCRγδ cells. RNA sequencing aligns M-ATO-derived populations with phenotypically identical primary thymocytes. M-ATOs initiated with Rag1(−/−) marrow produce the same differentiation block as seen in the endogenous thymus, and Notch signaling patterns in M-ATOs mirror primary thymopoiesis. M-ATOs initiated with defined hematopoietic stem cells (HSCs) and lymphoid progenitors from marrow and thymus generate each of the downstream differentiation stages, allowing the kinetics of T cell differentiation to be tracked. Remarkably, single HSCs deposited into each M-ATO generate the complete trajectory of T cell differentiation, producing diverse TCR repertoires across clones that largely match endogenous thymus. M-ATOs represent a highly reproducible and efficient experimental platform for the interrogation of clonal thymopoiesis from HSCs.