Ryu Kyung Ha, Cho Su Jin, Jung Yoon Jae, Seoh Ju Young, Kie Jeong Hae, Koh Sang Hyeok, Kang Hyoung Jin, Ahn Hyo Seop, Shin Hee Young
Department of Pediatrics, Ewha Womans University College of Medicine, Ewha Medical Research Center, Seoul, Korea.
Int J Hematol. 2004 Oct;80(3):281-6. doi: 10.1532/ijh97.a10406.
Dendritic cells (DCs) are the most potent antigen-presenting cells in terms of initiating primary T-cell-dependent immune responses. We devised a 2-step culture method for obtaining sufficient numbers of functional DCs from umbilical cord blood (CB) CD34+ cells. In the first step, CB CD34+ cells were expanded by stimulation with early-acting cytokines such as stem cell factor (SCF), flt3 ligand (FL), and thrombopoietin (TPO) to amplify the hematopoietic progenitor cells. In the second step, granulocyte-macrophage colony-stimulating factor and interleukin 4 were added, and incubation was continued for another 5 days to induce differentiation of the expanded cells into DCs. During the first step of culturing with TPO, SCF, and FL, the total numbers of nucleated cells gradually increased, peaking at 4 weeks (245.3-fold). During the second step, expression of CD1a, CD83, and CD86 increased. Electron microscopic findings showed that these cells had cytosolic expansion to form dendrites and major histocompatibility complex class II compartments, which are characteristic of DCs. Functional analyses revealed that these cells had phagocytic activity and were capable of stimulating allogeneic T-cells in vitro.
树突状细胞（DCs）是启动原发性T细胞依赖性免疫反应方面最有效的抗原呈递细胞。我们设计了一种两步培养方法，用于从脐带血（CB）CD34+细胞中获得足够数量的功能性DCs。第一步，通过用早期作用的细胞因子如干细胞因子（SCF）、fms样酪氨酸激酶3配体（FL）和血小板生成素（TPO）刺激来扩增CB CD34+细胞，以扩增造血祖细胞。第二步，加入粒细胞-巨噬细胞集落刺激因子和白细胞介素4，并继续培养5天，以诱导扩增的细胞分化为DCs。在用TPO、SCF和FL培养的第一步中，有核细胞总数逐渐增加，在4周时达到峰值（245.3倍）。在第二步中，CD1a、CD83和CD86的表达增加。电子显微镜检查结果显示，这些细胞具有胞质扩张以形成树突和主要组织相容性复合体II类区室，这是DCs的特征。功能分析表明，这些细胞具有吞噬活性，并且能够在体外刺激同种异体T细胞。
