Distinct subsets of dendritic cells resembling dermal DCs can be generated in vitro from monocytes, in the presence of different serum supplements
Distinct subsets of dendritic cells resembling dermal DCs can be generated in vitro from monocytes, in the presence of different serum supplements
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DOI：
10.1016/s0022-1759(00)00147-2
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发表时间：
2000-04-21
影响因子：
2.2
通讯作者：
Rigal, D
中科院分区：
文献类型：
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作者：
Duperrier, K;Eljaafari, A;Rigal, D
我们最近证明，只要细胞外 pH 值保持在血浆值，在高浓度人血清 (HS) 存在的情况下，单核细胞可以生成树突状细胞 (DC)。由于单核细胞来源的 DC (Mo-DC) 也可以在胎牛血清 (FCS) 或无血清培养基存在下生成，因此我们研究了这些不同的培养补充剂是否会影响 DC 的生成。为此，将纯化的单核细胞与 GM-CSF 加 IL-4 一起培养 6 天，并在 HS、自体血浆 (AP)、FCS 或无血清培养基 X-VIVO 20 存在的情况下进一步暴露于 TNF-α 2 天。我们的结果表明，通过 CD83 和 CD86 上调、葡聚糖-FITC 摄取、同种异体 MLR 测定和自体反应的诱导进行评估，在 HS 或 AP 存在的情况下可以重复获得功能成熟的 DC 的良好产量。有趣的是，血清对 DC 生成的影响可能不仅是定量的，而且是定性的，因为 (i) 大多数 HS 或 AP 培养的 DC 表达 CD83，CD1a 水平非常弱，而 FCS 或 X-VIVO 中培养的 CD83 + DC 大部分为 CD1++； (ii) HS 和 AP 培养的 DC 比 FCS 或 X-VIVO 培养的 DC 更具颗粒性和异质性，并且 (iii) 通过电子显微镜评估，在 HS 或 AP 培养的 DC 中优先观察到伯贝克样颗粒的存在。这些不同的细胞类似于真皮 DC (DDC)，这一点得到了观察结果的进一步支持：大多数细胞在不存在 Lag 抗原的情况下展示胞质内 FXIIIa，并以非常低的水平表达 E-钙粘蛋白。总而言之，我们的结果表明，根据培养条件，从相同的单核细胞群体开始，可以产生不同的 DC 子集。因此，HS或AP有利于生成类似于活化的真皮DC的完全成熟的DC，而FCS或X-VIVO优先导致不太成熟的CD1a+ + 真皮样DC的生成。 (C) 2000 Elsevier Science B.V. 保留所有权利。
We recently demonstrated that dendritic cells (DCs) can be generated from monocytes in the presence of high concentrations of human serum (HS), provided the extra-cellular pH is maintained at plasma values. Because monocyte-derived DCs (Mo-DCs) can also be generated in the presence of fetal calf serum (FCS) or serum-free medium, we have investigated whether these different culture supplements influence DC generation. With this aim, purified monocytes were cultured with GM-CSF plus IL-4 for 6 days and were further exposed to TNF-alpha for 2 additional days, in the presence of HS, autologous plasma (AP), FCS, or X-VIVO 20, a serum-free medium. Our results show that good yields of functionally mature DCs can reproducibly be obtained in the presence of HS or AP, as assessed by CD83 and CD86 up-regulation, dextran-FITC uptake, allogeneic MLR assays and the induction of an autologous response. Interestingly, the effect of serum on DC generation was probably not only quantitative, but also qualitative, since (i) the majority of HS- or AP-cultured DCs expressed CD83 with very weak levels of CD1a, whereas CD83 + DCs cultured in FCS or X-VIVO were mostly CD1+ +; (ii) HS- and AP-cultured DCs were much more granular and heterogeneous than FCS- or X-VIVO-cultured DCs, and (iii) the presence of Birbeck-like granules was preferentially observed in HS- or AP-cultured DCs, as assessed by electron microscopy. That these different cells resemble dermal DCs (DDCs) was further supported by the observations that most of the cells displayed intracytoplasmic FXIIIa in the absence of Lag antigen, and expressed E-cadherin at very low levels. Altogether, our results indicate that starting from the same monocytic population, different subsets of DCs can be generated, depending on the culture conditions. Thus, HS or AP favors the generation of fully mature DCs that resemble activated dermal DCs, whereas FCS, or X-VIVO preferentially leads to the generation of less mature CD1a+ + dermal-like DCs. (C) 2000 Elsevier Science B.V. All rights reserved.