HDFa (human dermal fibroblasts) are used as cellular models for EMF exposure. To ensure reproducible in vitro experiments, comparable proliferation and differentiation cell conditions must exist, and different donors, passage numbers, culture time, and growth media must be considered. In this study, the authors cultured fibroblasts in DMEM or 106 medium. Growth curves, vitality, morphology, and gene expression of genes coding for proliferation (PCNA, CDKN2A, CDKN1A, SFN), differentiation (PDGFRA, TGM2, ACTA2, PDPN, NTN1, MGP, PPP1R14), and SFN target genes (TP63, MMP1, MMP3) were examined in both media and passage numbers 3-4, 5-6 and >6. At passages 3-4, proliferating cells can be observed in both media. While cells cultured in DMEM proliferate over the passages, from passage 5, cells in 106 medium persisted around the seeded number. TGM2 down-regulation over all passages in both media and cells morphology suggest papillary-type fibroblasts. Downregulation of SFN (negative regulator of mitotic translation and cell differentiation) coincided with proliferating fibroblasts over all examined conditions. Downstream SFN target genes in proliferating cells appeared upregulated (TP63) and downregulated (MMP1/MMP3), suggestive for a status characterized by increased stemnesses (upregulated TP63) and wound healing capacity (downregulated MMP1, MMP3). Resting cells (SFN control values) were associated with control values of TP63 and MMP1/MMP3 expression, suggesting a reduced stemness and wound healing capacity. In conclusion, a set of markers related to proliferation (SFN), differentiation (TGM2), stemnesses (TP63), and wound healing (MMP1/MMP3) allow a culture characterization so that cells under two different conditions can be exposed, thus enabling reproducible EMF experiments or experiments with other exposures.

中文翻译：

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表征人类原代皮肤成纤维细胞，以确保在可比的细胞培养条件下进行EMF暴露实验。

HDFa（人类皮肤成纤维细胞）用作EMF暴露的细胞模型。为了确保可重复的体外实验，必须存在可比的增殖和分化细胞条件，并且必须考虑不同的供体，传代次数，培养时间和生长培养基。在这项研究中，作者在DMEM或106培养基中培养了成纤维细胞。编码增生（PCNA，CDKN2A，CDKN1A，SFN），分化（PDGFRA，TGM2，ACTA2，PDPN，NTN1，MGP，PPP1R14）和SFN目标基因（TP63，MMP1）的基因的生长曲线，活力，形态和基因表达（MMP3），分别以媒体号和3-4、5-6和> 6进行了检查。在第3-4代，可以在两种培养基中观察到增殖细胞。虽然在DMEM中培养的细胞在传代中增殖，但从传代5开始，在106个培养基中的细胞在种子数目附近持续存在。TGM2在培养基和细胞形态的所有传代中均下调，提示乳头状成纤维细胞。在所有检查条件下，SFN（有丝分裂翻译和细胞分化的负调节剂）的下调与增殖的成纤维细胞相吻合。增殖细胞中的下游SFN靶基因出现上调（TP63）和下调（MMP1 / MMP3），提示以干细胞增加（TP63上调）和伤口愈合能力（下调MMP1，MMP3）为特征的状态。静止细胞（SFN对照值）与TP63和MMP1 / MMP3表达的对照值相关，提示干细胞减少和伤口愈合能力下降。总之，一组与增殖（SFN），分化（TGM2），茎干（TP63），