Generation of T Cells from Human Embryonic Stem Cells.
Generation of T Cells from Human Embryonic Stem Cells.
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从人类胚胎干细胞生成 T 细胞。
DOI：
10.1182/blood.v112.11.1527.1527
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发表时间：
2008
期刊：
影响因子：
20.3
通讯作者：
B. Vandekerckhove
中科院分区：
文献类型：
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作者：
F. Timmermans;Imke Velghe;L. Walleghem;M. Smedt;S. V. Coppernolle;G. Leclercq;T. Taghon;H. Moore;A. Langerak;T. Kerre;J. Plum;B. Vandekerckhove
背景资料：人胚胎干细胞（human embryonic stem cells，hESC）来源于早期囊胚，其特征在于具有自我更新和产生分化的功能细胞类型的能力。hESC研究领域的主要挑战之一是建立一个培养系统，使hESC沿着特定的谱系命运发展。迄今为止，报告造血发育的研究尚未提供关于hESC体外分化为T谱系细胞的能力的证据。的选材及
方法：在所有实验中使用hESC系H1（美国国立卫生研究院[NIH]代码：WA 01），Wisconson，麦迪逊，美国）（第30-60代）。如前所述，hESC系在MEF上保持未分化状态。OP 9细胞和表达高水平Notch配体δ样1（OP 9-DLL 1，来自J.C. Zuniga-Pflucker，University of多伦多，加拿大），如前所述在含20%FCS的MEM-α中培养。
结果如下：我们的数据表明，T细胞可以在体外产生从hESC在一个强大的和高度可重复的方式使用hESC的顺序暴露于鼠OP 9细胞系和OP 9-DLL 1。在OP 9基质层上，产生了CD 34 highCD 43 dim造血前体群体，其局限于血管样结构，使人联想到体内胚胎发育期间出现的血岛。当暴露于0 P9-DLL 1单层时，该前体群体变成T谱系定型，依次经过CD 34 + CD 7+表型、CD 4 + CD 8+双阳性中间阶段，并最终分化成成熟T细胞。产生多克隆T细胞，细胞受体（TCR）α-β和TCR γ-δ，其基于增殖能力和TCR交联后细胞因子的产生而起作用。
结论：我们表明，成熟的和功能性的T细胞可以从人胚胎干细胞使用良好定义的体外条件。该方案与最近描述的诱导多能细胞相结合，可能在肿瘤免疫学中找到临床适用性。
Background: Human embryonic stem cells (hESC) are derived from early stage blastocysts and are characterized by the ability to both self-renew and to generate differentiated functional cell types. One of the major challenges in the field of hESC research, is to set up a culture system that drives hESC down a particular lineage fate. To date, studies reporting hematopoietic development have not provided evidence on the differentiation capacity of hESC into T lineage cells in vitro. Material and
Methods: hESC line H1 (National Institutes of Health [NIH] code: WA01), Wisconson, Madison, USA) was used (Passage 30–60) in all experiments. The hESC line was kept in an undifferentiated state on MEFs as previously described. OP9 cells and OP9 cells that express high levels of the Notch ligand Delta-like 1 (OP9-DLL1, a gift from J. C. Zuniga-Pflucker, University of Toronto, Canada) were cultured as previously described in MEM-α with 20 % FCS.
Results: Our data show that T cells can be generated in vitro from hESC in a robust and highly reproducible manner using the sequential exposure of hESC to the murine OP9 cell line and OP9-DLL1. On OP9 stromal layers, a CD34highCD43dim hematopoietic precursor population is generated that is confined to vascular-like structures, reminiscent of blood islands that emerge during in vivo embryonic development. This precursor population becomes T lineage committed when exposed to OP9-DLL1 monolayers, passing sequentially through a CD34+CD7+ phenotype, a CD4+CD8+ double positive intermediate stage and eventually differentiates into a mature T cells. Polyclonal T cells are generated, cell receptor (TCR) alpha-beta and TCRgamma-delta which are functional based on proliferative capacity and production of cytokines after TCR crosslinking.
Conclusion: We show that mature and functional T cells can be generated from hESC using well defined in vitro conditions. This protocol in combination with the recently described induced pluripotent cells may find clinical applicability in tumor immunology.