DENDRITIC CELLS FROM HUMAN EMBRYONIC STEM CELLS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 目的：探讨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hESCs向髓样DCs高效分化的体外系统。与0 P9基质细胞共培养的hESC诱导其分化为髓样祖细胞。 为了产生DC，我们在GM-CSF存在下扩增了hESC衍生的髓样祖细胞。 产生的髓系祖细胞具有CD 4 + CD 11b + CD 11 c + CD 16 + CD 123 lowHLA-DR-表型并表达髓过氧化物酶。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、表达高水平的MHC I类和II类分子、CD 1a、CD 11 c、CD 80、CD 86、DC-SIGN和CD 40的细胞。hEC-DC能够摄取和加工抗原，如DQ卵白蛋白测定所确定的，触发MLR中的幼稚T细胞，并通过MHC I类途径将抗原呈递给特异性T细胞克隆。目前项目的总体目标是应用所描述的模型来鉴定髓样DC前体和调节其扩增的通路。 这项研究使用了WNPRC干细胞资源。 出版物： Choi KD，Vodyanik MA，Slukvin II.多能干细胞体外扩增和诱导分化为成熟人骨髓单核细胞衍生的lin-CD 34 + CD 43 + CD 45+祖细胞。临床研究杂志。2009年9月;119（9）：2818-29。电子版2009年8月10日。PMID：19726877。

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 in vitro system for efficient differentiation of hESCs into myeloid DCs. hESCs cocultured 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. Generated myeloid progenitors 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 current 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. PUBLICATION: Choi KD, Vodyanik MA, Slukvin II. Generation of mature human myelomonocytic cells through expansion and differentiation of pluripotent stem cellderived lin-CD34+CD43+CD45+ progenitors. Journal of Clinical Investigation. Sept 2009;119(9):2818-29. Epub 2009 Aug 10.PMID: 19726877.