DENDRITIC CELLS FROM HUMAN EMBRYONIC STEM CELLS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：促进人胚胎干细胞来源的树突状细胞在血液病治疗中的临床应用。 我们建立了人胚胎干细胞高效分化为髓系树突状细胞的体外系统。我们将人胚胎干细胞与OP9基质细胞共培养，诱导其分化为髓系祖细胞。为了产生DC，我们在GM-CSF的存在下扩增了hESC来源的髓系祖细胞。我们培养出的髓系祖细胞具有CD4+CD11b+CD11c+CD16+CD123低HLA-DR表型，并表达髓过氧化物酶。髓系细胞在GM-CSF和IL-4的无血清培养条件下，可产生具有典型树突状形态的细胞，高水平表达MHC I、II类分子CD1a、CD11c、CD80、CD86、DC-SIGN和CD40。DQ卵白蛋白检测结果表明，HEC-DC具有摄取和处理抗原的能力，触发MLR中的NA‘ve T细胞，并通过MHC-I类途径将抗原递呈给特定的T细胞克隆。这个正在进行的项目的总体目标是应用所描述的模型来识别髓系树突状细胞前体和调节其扩张的途径。 这项研究使用了WNPRC干细胞资源和联邦批准的HES细胞系。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To advance human embryonic stem cell-derived dendritic cells towards clinical application for treatment of blood disease. We established an in vitro system for efficient differentiation of hESCs into myeloid DCs. We cocultured hESCs with OP9 stromal cells to induce their differentiation into myeloid progenitors. To produce DCs, we expanded hESC-derived myeloid progenitors in presence of GM-CSF. We generated myeloid progenitors that had CD4+CD11b+CD11c+ CD16+ CD123lowHLA-DR- phenotype and expressed myeloperoxidase. Further culture of myeloid cells in serum free media with GM-CSF and IL-4 generated cells that had typical dendritic morphology, expressed high level of MHC class I and II molecules, CD1a, CD11c, CD80, CD86, DC-SIGN and CD40. The hEC-DCs were capable of uptaking and processing antigen, as determined by DQ ovalbumin assay, triggering na¿ve T cells in MLR and presenting antigen to specific T cell clones through MHC class I pathway. The overall goal of this ongoing project is to apply the described model for identification of myeloid DC precursors and pathways regulating their expansion. This research used WNPRC stem cell resources and federally approved hES cell lines.