Background and aims

Immortalized cell lines have been long used as substitute for ex vivo murine and human material, but exhibit features that are not found in healthy tissue. True human dendritic cells (DC) cannot be cultured or passaged as opposed to immortalized cell lines. Research in the fields of immunogenic responses and immunotolerance in DCs has increased over the last decade. Autophagy has gained interest in these fields as well, and has been researched extensively in many other cell types as well. Here we have studied the applicability of cell line-derived dendritic cell-like cells of six myeloid cell lines aimed at research focussed on autophagy.

Methods

Six myeloid leukaemia cell lines were differentiated towards cell line-derived dendritic cell-like cells (cd-DC) using GM-CSF, IL-4, Ionomycine and PMA: HL60, KG1, MM6, MV-4-11, THP1 and U937. Autophagy was modulated using Rapamycin, Bafilomycin A1 and 3MA. Cell lines were genotyped for autophagy-related SNPs using RFLP. Marker expression was determined with FACS analysis and cytokine profiles were determined using Human Cytometric Bead Assay. Antigen uptake was assessed using Fluoresbrite microspheres.

Results and discussion

All researched cell lines harboured SNPs in the autophagy pathways. MM6 and THP1 derived cd-DCs resembled monocyte-derived DCs (moDC) most closely in marker expression, cytokine profiles and autophagy response. The HL60 and U937 cell lines proved least suitable for autophagy-related dendritic cell research.

Conclusion

The genetic background of cell lines should be taken into account upon studying (the effects of) autophagy in any cell line. Although none of the studied cell lines recapitulate the full spectrum of DC characteristics, MM6 and THP1 derived cd-DCs are most suitable for autophagy-related research in dendritic cells.

中文翻译：

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不同细胞系来源的树突状细胞样细胞在自噬研究中的适用性

背景和目标

永生化细胞系长期以来一直被用作离体小鼠和人类材料的替代品，但表现出健康组织中没有的特征。与永生化细胞系不同，真正的人类树突状细胞 (DC) 无法培养或传代。在过去十年中，DC 的免疫原性反应和免疫耐受领域的研究有所增加。自噬也引起了这些领域的兴趣，并且在许多其他细胞类型中也得到了广泛的研究。在这里，我们研究了六种骨髓细胞系的细胞系衍生的树突状细胞样细胞的适用性，旨在研究专注于自噬。

方法

使用 GM-CSF、IL-4、离子霉素和 PMA 将六种髓系白血病细胞系分化为细胞系衍生的树突状细胞样细胞 (cd-DC)：HL60、KG1、MM6、MV-4-11、THP1 和 U937 . 使用雷帕霉素、巴弗洛霉素 A1 和 3MA 调节自噬。使用 RFLP 对细胞系进行自噬相关 SNP 的基因分型。标记表达通过 FACS 分析确定，细胞因子谱通过人细胞计数珠分析确定。使用 Fluoresbrite 微球评估抗原摄取。

结果和讨论

所有研究的细胞系在自噬途径中都含有 SNP。MM6 和 THP1 衍生的 cd-DCs 在标志物表达、细胞因子谱和自噬反应方面与单核细胞衍生的 DCs (moDC) 最相似。HL60 和 U937 细胞系被证明最不适合自噬相关的树突细胞研究。

结论

在研究任何细胞系中的自噬（影响）时，应考虑细胞系的遗传背景。尽管所研究的细胞系都没有概括 DC 特征的全谱，但 MM6 和 THP1 衍生的 cd-DC 最适合树突状细胞的自噬相关研究。