体外重现单个造血干细胞来源的鼠类胸腺发生。

In Vitro Recapitulation of Murine Thymopoiesis from Single Hematopoietic Stem Cells.

机构信息

Department of Pathology & Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles (UCLA), Los Angeles, CA, USA.

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.

出版信息

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

DOI:10.1016/j.celrep.2020.108320

PMID:33113379

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7727762/

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.

摘要

我们报告了一种无血清的 3D 小鼠人工胸腺类器官（M-ATO）系统，该系统模拟了正常的小鼠胸腺发生，产生了所有 T 细胞阶段，从早期胸腺祖细胞到功能性单阳性（CD8SP 和 CD4SP）TCRαβ和 TCRγδ细胞。RNA 测序将 M-ATO 衍生的群体与表型相同的原代胸腺细胞进行了匹配。用 Rag1 骨髓启动的 M-ATOs 产生了与内源性胸腺中相同的分化阻断，并且 M-ATOs 中的 Notch 信号模式反映了原代胸腺发生。用骨髓和胸腺中的定义造血干细胞（HSCs）和淋巴祖细胞启动的 M-ATOs 产生了下游分化阶段中的每一个，从而可以追踪 T 细胞分化的动力学。值得注意的是，每个 M-ATO 中沉积的单个 HSCs 产生了 T 细胞分化的完整轨迹，在克隆中产生了多样化的 TCR 库，这些库在很大程度上与内源性胸腺相匹配。M-ATOs 代表了一种高度可重复和高效的实验平台，可用于从 HSCs 中探究克隆性胸腺发生。