Comparison of adherent and non-adherent cell phenotypes of breast carcinoma cultures

Introduction. Primary cell cultures of breast carcinoma (BC) are used as a model for study the processes of intratumoral heterogeneity, drug resistance and various molecular and biological processes. One of the variants of primary culture is non-adhesive cell culture in the form of spheroids – mammospheres. There is evidence that cells isolated from mammospheres have mesenchymal features. However, the acquisition of mesenchymal features in mammospheres derived from tumors correlates with suppression of estrogen receptor expression and, therefore, with resistance to therapy against hormone-dependent tumors.

The aim of the study was to determine the effect of the method of culturing BC cells on the ability of cells to preserve the epithelial phenotype.

Materials and methods A comparative analysis of two BC culture samples examined over three passages using different methods of cultivation was performed. Staining for morphological evaluation was carried out according to Pappenheim. Determination of belonging to epitheliocytes was performed using anti-Pan Keratin (AE1/AE3/ PCK26) Primary Antibody (Roche diagnostics, USA).

Results Three passages (P2, P3, P4) of non-adherent and three passages of adherent cells were taken for culture analysis. During the study of these BC culture variants, the morphological features of the cells in each passage were determined and it was found that regardless of the technique chosen during cultivation, the cells of the cultures under study retained the epithelial phenotype. However, the study of the adhesive culture revealed a higher percentage of cells showing the epithelial phenotype compared to the cells of the non-adhesive culture.

Discussion In the course of our study, six morphological groups of cells obtained during the creation of a primary culture of a BC sample were identified, among which differences in the preservation of the epithelial nature throughout the cultivation were manifested.

Conclusion The choice of cultivation technique influences the ability of cells to retain the epithelial phenotype for three passages. In the second passage, the level of pancytokeratin expression in the adhesive culture is significantly differed from that in the non-adhesive culture. By the fourth passage there is a sharp decrease in the number of epithelial cells.

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