In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells

Amélie Montel-Hagen; Victoria Sun; David Casero; Steven Tsai; Alexandre Zampieri; Nicholas Jackson; Suwen Li; Shawn Lopez; Yuhua Zhu; Brent Chick; Chongbin He; Stéphanie C de Barros; Christopher S Seet; Gay M Crooks

doi:10.1016/j.celrep.2020.108320

In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells

Cell Rep. 2020 Oct 27;33(4):108320. doi: 10.1016/j.celrep.2020.108320.

Authors

Affiliations

¹ Department of Pathology & Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles (UCLA), Los Angeles, CA, USA.
² Department of Pathology & Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles (UCLA), Los Angeles, CA, USA; Molecular Biology Interdepartmental Program, UCLA, Los Angeles, CA, USA.
³ Division of Hematology-Oncology, Department of Medicine, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA.
⁴ Department of Pathology & Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles (UCLA), Los Angeles, CA, USA; Department of Molecular and Medical Pharmacology, UCLA, Los Angeles, CA, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, UCLA, Los Angeles, CA, USA.
⁵ Division of Hematology-Oncology, Department of Medicine, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, UCLA, Los Angeles, CA, USA; Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA, USA.
⁶ Department of Pathology & Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles (UCLA), Los Angeles, CA, USA; Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, UCLA, Los Angeles, CA, USA; Jonsson Comprehensive Cancer Center, UCLA, Los Angeles, CA, USA; Division of Pediatric Hematology-Oncology, Department of Pediatrics, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA. Electronic address: gcrooks@mednet.ucla.edu.

Abstract

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.

Keywords: Hematopoietic stem and progenitor cells; T-cells; Thymopoiesis; in vitro T-cell development; single cell; thymic organoids.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation
Hematopoietic Stem Cells / cytology
Hematopoietic Stem Cells / metabolism*
Mice
Thymus Gland / physiology*

Abstract

Publication types

MeSH terms

Grants and funding