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Role of geneticin in isolation and culturing of skin melanocytes and melanoma cells

Postępy Higieny i Medycyny Doświadczalnej
Volume 77 (2023): Issue 1 (January 2023)
By:
Open Access
|Sep 2023

Figures & Tables

Figure 1.

Schematic presentation of the analysis of G418 influence on cells’ metabolic activity
Schematic presentation of the analysis of G418 influence on cells’ metabolic activity

Figure 2.

Viability of normal human skin cells, cultured in M254 culture medium, after cell culture stimulation with different concentrations of geneticin for 24 hours (A) and 72 hours (B)
Viability of normal human skin cells, cultured in M254 culture medium, after cell culture stimulation with different concentrations of geneticin for 24 hours (A) and 72 hours (B)

Figure 3.

The viability of primary keratinocytes measured 24 h (A) and 72 h (B) after cell culture stimulation with increasing concentrations of geneticin, measured by the means of MTT assay. All absorbance values were normalized versus control keratinocytes obtained after 24 hours of cell culture
The viability of primary keratinocytes measured 24 h (A) and 72 h (B) after cell culture stimulation with increasing concentrations of geneticin, measured by the means of MTT assay. All absorbance values were normalized versus control keratinocytes obtained after 24 hours of cell culture

Figure 4.

The viability of primary fibroblasts measured 24 h (A) and 72 h (B) after cell culture stimulation with increasing concentrations of geneticin measured by the means of MTT assay. All absorbance values were normalized versus control fibroblasts obtained after 24 hours of cell culture
The viability of primary fibroblasts measured 24 h (A) and 72 h (B) after cell culture stimulation with increasing concentrations of geneticin measured by the means of MTT assay. All absorbance values were normalized versus control fibroblasts obtained after 24 hours of cell culture

Figure 5.

The proliferation rates of primary fibroblasts cultured in different media supplemented with geneticin (0.05–1 mg/ml) measured using the BrdU assay (A). The quantitative analysis of apoptosis and necrosis of fibroblasts treated with geneticin with staurosporine/etoposide as positive control samples (K(+)), respectively (B)
The proliferation rates of primary fibroblasts cultured in different media supplemented with geneticin (0.05–1 mg/ml) measured using the BrdU assay (A). The quantitative analysis of apoptosis and necrosis of fibroblasts treated with geneticin with staurosporine/etoposide as positive control samples (K(+)), respectively (B)

Figure 6.

Immunofluorescence staining of melanocytes, keratinocytes and fibroblasts cultured in medium M254 for 72 hours after 0.05 mg/mL geneticin treatment. The nuclei of the cells were stained blue with DAPI. TYR and NG2 are markers of melanocytes, while COL1 and KRT14 are markers of fibroblasts and keratinocytes, respectively
Immunofluorescence staining of melanocytes, keratinocytes and fibroblasts cultured in medium M254 for 72 hours after 0.05 mg/mL geneticin treatment. The nuclei of the cells were stained blue with DAPI. TYR and NG2 are markers of melanocytes, while COL1 and KRT14 are markers of fibroblasts and keratinocytes, respectively

Figure 7.

The MTT results of the influence of geneticin on the primary human metastatic melanoma cell lines (WCCm1, WCCm7, WCCd9) and commercial cell lines of metastatic melanoma (A375, G-361 and MeWo) observed 24 hours (A) and 72 hours (B) after cell stimulation in M254 culture medium
The MTT results of the influence of geneticin on the primary human metastatic melanoma cell lines (WCCm1, WCCm7, WCCd9) and commercial cell lines of metastatic melanoma (A375, G-361 and MeWo) observed 24 hours (A) and 72 hours (B) after cell stimulation in M254 culture medium

Figure 8.

Flow of information through the selected literature about concentrations of geneticin used for culture of pure melanocytes
Flow of information through the selected literature about concentrations of geneticin used for culture of pure melanocytes

Percentage of particular cells in co-cultures of melanocytes, fibroblasts, and keratinocytes in different culture media for 72 hours following treatment with 0_05 or 0_1 mg/mL geneticin

Culture mediaCell typeControlGeneticin (0.05 mg/mL)Geneticin (0.1 mg/mL)
DMEM + 10% FBSMelanocytes15%29%37%
Fibroblasts67%66%60%
Keratinocytes18%5%3%
RPMI + 10% FBSMelanocytes26%40%66%
Fibroblasts60%55%30%
Keratinocytes14%5%4%
M254Melanocytes27%97%98%
Fibroblasts40%2%1%
Keratinocytes33%1%1%
DOI: https://doi.org/10.2478/ahem-2023-0014 | Journal eISSN: 1732-2693
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Language: English
Page range: 72 - 81
Submitted on: Mar 12, 2023
Accepted on: Jul 6, 2023
Published on: Sep 14, 2023
Published by: Hirszfeld Institute of Immunology and Experimental Therapy
In partnership with: Paradigm Publishing Services
Publication frequency: 1 times per year
Keywords:
melanocytes,
fibroblasts,
keratinocytes,
melanoma,
geneticin,
cell culture contamination
Related subjects:
Life sciences,
Molecular biology,
Microbiology and virology,
Medicine,
Basic medical science,
Immunology

© 2023 Aneta Ścieżyńska, Anna Sobiepanek, Marta Soszyńska, Krzysztof Łuszczyński, Marcin Radziszewski, Iryna Levkovych, Natalia Krześniak, Beata Orzechowska, Anna Lutyńska, Jacek Malejczyk, published by Hirszfeld Institute of Immunology and Experimental Therapy
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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