Ex vivo expansion of canine dendritic cells from CD34+ bone marrow progenitor cells
Ex vivo expansion of canine dendritic cells from CD34+ bone marrow progenitor cells
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DOI：
10.1097/00007890-200011270-00007
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发表时间：
2000-11-27
期刊：
影响因子：
6.2
通讯作者：
Nash, RA
中科院分区：
文献类型：
--
作者：
Hägglund, HG;McSweeney, PA;Nash, RA
背景本研究的目的是体外扩增犬树突状细胞，并确定其表型和功能特性。在Iscove改良培养基中培养CD34（+）选择的细胞和CD34（+）去除的犬骨髓（BM）细胞14天。加入培养物的细胞因子包括人粒细胞/巨噬细胞集落刺激因子5 ng/ml、hFlt3配体200 ng/ml和人肿瘤坏死因子-α 10 ng/ml。培养细胞和纯化的亚群通过流式细胞仪分析、电子显微镜和同种异体混合淋巴细胞反应在第14天评估细胞表面抗原表达、形态和功能。两个主要细胞。DR ++（亮）/CD14（--）和DR+（暗）/CD14（+）。体外扩增的CD34+选择性细胞与培养的CD34+耗竭细胞和单核细胞相比，显示出增加的同种异体刺激活性。相比之下，从CD34（+）耗尽的细胞的离体扩增是不成功的。从体外扩增的CD 34（+）选择的骨髓中分选细胞以富集DR ++/CD 14（--）细胞后，与分选的DR +/CD 14（+）细胞相比，观察到同种异体刺激活性增加42倍（中位数）（P = 0.02）。具有树突状细胞样表型和功能的细胞可以从犬CD 34（+）选择的骨髓细胞中培养。未来的研究将探讨这些细胞在犬造血干细胞移植模型中的植入、移植物抗宿主反应和移植物-宿主耐受中的作用。
Background. The aims of this study were to ex vivo expand canine dendritic cells and determine their phenotype and functional characteristics.Methods. CD34(+)-selected cells and CD34(+)-depleted canine bone marrow (BM) cells were cultured in Iscove's modified medium for 14 days. Cytokines added to the cultures included human granylocyte/ macrophage colony-stimulating factor 5 ng/ml, hFlt3 ligand 200 ng/ml, and human tumor necrosis factor-alpha 10 ng/ml. Cultured cells and purified subpopulations were assessed for cell surface antigen expression, morphology, and function by flow cytometric analysis, electron microscopy, and an allogeneic mixed lymphocyte reaction at day 14.Results. Two main cell. populations were identified, DR++(bright)/CD14(--) and DR+(dim)/CD14(+). Ex vivo expanded CD34+-selected cells showed increased allostimulatory activity compared to both cultured CD34(+)-depleted cells and mononuclear cells. In contrast, ex vivo expansion from CD34(+)-depleted cells was unsuccessful. After sorting cells from the ex vivo expanded CD34(+)-selected bone marrow to enrich for DR++/CD14(--) cells, a 42-fold increase (median) of allostimulatory activity was observed as compared with sorted DR+/CD14(+) cells (P=0.02).Conclusions. Cells with dentric cell-like phenotypes and functions can be cultured from canine CD34(+)-selected bone marrow cells. Future studies will address the roles of these cells in engraftment, graft versus host reactions and graft-host tolerance in a canine hematogoietic stem cell transplantaton model.