Determination of the Epithelial Nature of Cells and the Level of Proliferative Activity of Ki‑67 in the Primary Culture of Breast Carcinoma

Abstract

Introduction. Despite the fact that the immunohistochemical method is the most widely used in clinical practice, its use in the study of cell cultures is impossible, therefore, expression of molecular markers at the cellular level, the immunocytochemical method, is used as an alternative method. The aim of the study was to study the possibility of primary cell culture to maintain the proliferative ability and epithelial nature of cells over several passages. Material and methods. The paper describes a case of obtaining a primary cell culture of breast carcinoma. A study of the Ki‑67 cell proliferation index and determination of the epithelial nature of the cells was carried out. Results. When determining the index of cell proliferation Ki‑67, there was a tendency to increase this index at the second passage. The lowest level was noted at the first passage and amounted to 8.9 % of stained cells. To determine the possibility of using the obtained culture after the cryopreservation procedure, a second study was performed for both proteins at the third passage. The data obtained do have statistically significant differences according to the Mann–Whitney test and are 27.3 % and 47.2 % for Ki‑67, 75 % and 81 % for cytokeratin expression, respectively. Discussion. The level of the Ki‑67 cell proliferation index corresponded to the level of the initial sample at the first passage, when the cell culture was received. The cells showed a high index during further replanting. This indicates a high level of proliferative processes. This may be due to the peculiarities of the receptor apparatus of cells. The cytokeratin level indicates the epithelial nature of the tumor. This should be taken into account in various kinds of molecular studies of epithelial cells in vitro. Conclusion. The primary culture of breast carcinoma during cultivation retains a predominantly epithelial nature for three passages and after cryopreservation. The cell proliferation index Ki‑67 changes during culture subcultures.