Directed differentiation of human embryonic stem cells into functional dendritic cells through the myeloid pathway
Directed differentiation of human embryonic stem cells into functional dendritic cells through the myeloid pathway
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DOI：
10.4049/jimmunol.176.5.2924
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发表时间：
2006-03-01
影响因子：
4.4
通讯作者：
Choi, Kyung-Dal
中科院分区：
文献类型：
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作者：
Slukvin, Igor I.;Vodyanik, Maxim A.;Choi, Kyung-Dal
我们建立了一个将人胚胎干（hES）细胞定向分化为骨髓树突状细胞（DC）的系统。第一步，我们通过将 hES 细胞与 OP9 基质细胞共培养来诱导造血分化，然后使用无饲养层培养系统与 GM-CSF 一起扩增骨髓细胞。骨髓细胞具有CD4(+)CD11b(+)CD11c(+)CD16(+)CD123(低)HLA-DR-表型，表达髓过氧化物酶，并包括一群M-CSFR+单核细胞谱系定型细胞。在含有 GM-CSF 和 IL-4 的无血清培养基中进一步培养骨髓细胞，产生具有典型树突状形态的细胞；表达高水平的 MHC I 类和 II 类分子、CD1a、CD11c、CD80、CD86、DC-SIGN 和 CD40；并且能够处理 Ag，触发 NILR 中的初始 T 细胞，并通过 MHC I 类途径将 Ag 呈递给特定的 T 细胞克隆。将 DC 与 A23187 钙离子载体孵育 48 小时，诱导成熟 DC 标记物 CD83 和肌成束蛋白的表达。 GM-CSF与IL-4的组合为DC分化提供了最佳条件。用GM-CSF和TNF-α获得的DC共表达高水平的CD14，并且MLR刺激能力低。这些数据清楚地表明，hES细胞可以用作造血和DC前体以及不同成熟阶段的DC的新颖且独特的来源，以解决DC发育和生物学的基本问题。此外，由于ES细胞可以无限制地扩增，因此它们可以被视为DC疫苗或DC介导的免疫耐受诱导的潜在可扩展细胞来源。
We have established a system for directed differentiation of human embryonic stem (hES) cells into myeloid dendritic cells (DCs). As a first step, we induced hemopoietic differentiation by coculture of hES cells with OP9 stromal cells, and then, expanded myeloid cells with GM-CSF using a feeder-free culture system. Myeloid cells had a CD4(+)CD11b(+)CD11c(+)CD16(+) CD123(low)HLA-DR- phenotype, expressed myeloperoxidase, and included a population of M-CSFR+ monocyte-lineage committed cells. Further culture of myeloid cells in serum-free medium with GM-CSF and IL-4 generated cells that had typical dendritic morphology; expressed high levels of MHC class I and II molecules, CD1a, CD11c, CD80, CD86, DC-SIGN, and CD40; and were capable of Ag processing, triggering naive T cells in NILR, and presenting Ags to specific T cell clones through the MHC class I pathway. Incubation of DCs with A23187 calcium ionophore for 48 h induced an expression of mature DC markers CD83 and fascin. The combination of GM-CSF with IL-4 provided the best conditions for DC differentiation. DCs obtained with GM-CSF and TNF-alpha coexpressed a high level of CD14, and had low stimulatory capacity in MLR. These data clearly demonstrate that hES cells can be used as a novel and unique source of hemopoietic and DC precursors as well as DCs at different stages of maturation to address essential questions of DC development and biology. In addition, because ES cells can be expanded without limit, they can be seen as a potential scalable source of cells for DC vaccines or DC-mediated induction of immune tolerance.