DL4-μbeads induce T cell lineage differentiation from stem cells in a stromal cell-free system.
DL4-μbeads induce T cell lineage differentiation from stem cells in a stromal cell-free system.
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DOI：
10.1038/s41467-021-25245-8
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发表时间：
2021-08-18
影响因子：
16.6
通讯作者：
Zúñiga-Pflücker JC
中科院分区：
文献类型：
--
作者：
Trotman-Grant AC;Mohtashami M;De Sousa Casal J;Martinez EC;Lee D;Teichman S;Brauer PM;Han J;Anderson MK;Zúñiga-Pflücker JC
T细胞是免疫系统的关键效应器，可以作为再生医学和癌症免疫治疗的治疗手段。该领域尚未遇到的挑战是开发一种临床相关系统，该系统易于扩展，可以从造血干细胞/祖细胞（HSPCs）中产生大量t系细胞。在这里，我们报告了一种无基质细胞、基于微珠的方法，该方法支持人类祖T细胞（proT）和T谱系细胞的高效体外发育，这些细胞来自脐带血、gcsf动员的外周血和多能干细胞（PSCs）中的CD34+细胞。DL4 μ珠与淋巴细胞因子一起诱导CD34+细胞向CD34+CD7+CD5+ proT细胞向CD3+αβ T细胞的有序分化。人类psc衍生的proT细胞的单细胞RNA测序揭示了类似于胚胎和胎儿胸腺中发现的最早的胸腺细胞的转录谱。此外，CD34+CD7+ proT细胞在免疫缺陷小鼠体内的过继转移显示了有效的胸腺植入和外周T细胞的功能成熟。DL4 μbeads提供了一个简单而强大的平台来研究人类T细胞的发育，并促进了可再生资源工程T细胞疗法的开发。从干细胞中提取的T细胞可以用于再生医学和癌症免疫治疗，但目前的技术限制了T细胞的生产和转化。在这里，作者提出了一种无血清、无基质细胞的dll4包被微珠方法，用于从多种干细胞来源大规模生产t系细胞。
T cells are pivotal effectors of the immune system and can be harnessed as therapeutics for regenerative medicine and cancer immunotherapy. An unmet challenge in the field is the development of a clinically relevant system that is readily scalable to generate large numbers of T-lineage cells from hematopoietic stem/progenitor cells (HSPCs). Here, we report a stromal cell-free, microbead-based approach that supports the efficient in vitro development of both human progenitor T (proT) cells and T-lineage cells from CD34+cells sourced from cord blood, GCSF-mobilized peripheral blood, and pluripotent stem cells (PSCs). DL4-μbeads, along with lymphopoietic cytokines, induce an ordered sequence of differentiation from CD34+ cells to CD34+CD7+CD5+ proT cells to CD3+αβ T cells. Single-cell RNA sequencing of human PSC-derived proT cells reveals a transcriptional profile similar to the earliest thymocytes found in the embryonic and fetal thymus. Furthermore, the adoptive transfer of CD34+CD7+ proT cells into immunodeficient mice demonstrates efficient thymic engraftment and functional maturation of peripheral T cells. DL4-μbeads provide a simple and robust platform to both study human T cell development and facilitate the development of engineered T cell therapies from renewable sources. T cells derived from stem cells can be harnessed for regenerative medicine and cancer immunotherapy, but current technologies limit production and translation. Here, the authors present a serum-free, stromal-cell free DLL4-coated microbead method for the scalable production of T-lineage cells from multiple sources of stem cells.