keratinocyte basal media 2  (PromoCell)

Journal: Journal of Tissue Engineering

Article Title: An organotypic oral mucosal infection model to study host-pathogen interactions

doi: 10.1177/20417314231197310

Figure Lengend Snippet: OMMs display histological similarities to native oral mucosal tissue. Images of normal human oral mucosa (a and b) were obtained from: A histopathological image repository of normal epithelium of Oral Cavity and Oral Squamous Cell Carcinoma by Rahman et al. , under CC BY 4.0, images were re-oriented, cropped, labelled and post-fitted with a scale bar The two images were selected as they represented a similar field of view to the OMMs formed in this study. A representative sample of paraffin embedded, 4 μm sectioned, haematoxylin and eosin-stained oral mucosal tissue (images c and d) at day 19 of growth (100× magnification) were evaluated. Two images are selected to demonstrate the range of histological presentation of the OMMs and post-fitted with scale bars. Image scale was calculated using pixel size, number of pixels, width of the output image and objective magnification. Labels indicate the epithelial (keratinocyte-containing) and the lamina propria (fibroblast-containing) layers of the tissue.

Article Snippet: Once the models had contracted (~48 h), remaining media was aspirated and replaced with 500 μl of KGM: Keratinocyte gold basal media, Lonza, Slough, UK, supplemented with KGM gold bullet kit, Lonza (omitting Gentamicin/Amphotericin) and 2.4 μM CaCl 2 , below the transwell insert.

Techniques: Staining

Journal: Journal of Tissue Engineering

Article Title: An organotypic oral mucosal infection model to study host-pathogen interactions

doi: 10.1177/20417314231197310

Figure Lengend Snippet: Collagen and conditioned media differentially induce fibroblast and keratinocyte pro-inflammatory cytokines. IL-6 production by HaCaT (a), IL-6 production by HGF (b), IL-8 production by HaCaT (c) and IL-8 production by HGF (d), in response to collagen and supernatant (SN). Supernatant was pulse-chased to ensure that any IL-6 or IL-8 observed by ELISA was because of stimulation by the other incorporated cell-type. Data are from three independent experiments performed in triplicate. Significant differences in cytokine production were compared between the unstimulated control (media only) and stimulated groups. Data normality was assessed using the Shapiro-Wilk test for normality. Statistical significance was determined using a Kruskal-Wallis test followed by a Dunn’s multiple comparisons test. Statistical significance is indicated on the graph (* p < 0.05. ** p < 0.01. *** p < 0.001. **** p < 0.0001, ns = not significant). Error bars represent standard deviation.

Article Snippet: Once the models had contracted (~48 h), remaining media was aspirated and replaced with 500 μl of KGM: Keratinocyte gold basal media, Lonza, Slough, UK, supplemented with KGM gold bullet kit, Lonza (omitting Gentamicin/Amphotericin) and 2.4 μM CaCl 2 , below the transwell insert.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Standard Deviation