Skip to main content
. 2023 Jun 9;9(6):e17141. doi: 10.1016/j.heliyon.2023.e17141

Fig. 8.

Fig. 8

Pro- and anti-inflammatory cytokine expression by quantitative RT-PCR and immunoassay in statichBM-MSC-hSkM-PBMC co-culture. (a) hBM-MSCs were co-seeded with hSkMs in ratio 2:1 and PBMCs were culture in the upper chamber of transwell inserts. mRNA levels of pro-inflammatory (TNF, IL12A, and IL1B) and anti-inflammatory cytokines (IL10, TGFB1, and IL4) were assayed by qRT-PCR at day 7, 14 and 21 of culture. The relative quantification of each mRNA gene expression normalized to endogenous GAPDH (internal control) was calculated using the 2−ΔΔCt method and presented as fold change over hBM-MSCs at day 0, used as control. (b) Cytokine levels were also measured in culture medium supernatants at day 7, 14, and 21 by magnetic bead-based multiplex immunoassay. Data are reported as heatmap from blue (lowest value, 0 pg/mL of concentration) to red (highest value). Cytokines were hierarchical clustered based on expression pattern. Heatmap was made using Pheatmap and ComplexHeatmap packages in R Studio software (v. 2022.07.1 + 554; R Studio, Boston, MA, US). *p < 0.05; **p < 0.01; ****p < 0.0001.