Bulletin "Veterinary biotechnology"

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Veterynarna biotehnologija – Veterinary biotechnology, 2016, 29, 147-156 [in Ukrainian].

KOVPAK O.S., e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it.

National University of Life and Environmental Sciences of Ukraine

PHENOTYPIC AND MORPHOLOGICAL CHANGES OF MYOCARDIAL CELLS CULTURE DURING CULTIVATION

Introduction. The current practice of cell therapy implementation requires meticulous expert analysis of source material, careful detalization each of the stages of the using cells material and tracking the fate of the last one in vitro or in vivo. Therefore phenotyping of the cell culture seems to be relevant and timely.

Goal of the work. Investigate morphological and phenotypic changes of myocardial cells during their cultivation in vitro from the first to the fourth passage.

Materials and methods. Culture of the rats myocardial cells of the first to the fourth passages were used in this research. Control of phenotype changes were performed by detection of CD-markers (СD10, CD38, CD34, CD45, CD48, CD54, CD56, CD66e, CD96, CD227, CD326, рan keratin) and cardiomyocyte-specific protein – troponin I. Morphological changes of culture myocardial cells were evaluated visually.

Results of research and discussion. Our research revealed that the expression of cardiac muscle marker (СD10, CD95, CD 38, troponin I) decreased during cultivation. Meantime the intensity of markers expression, which characterized by hematopoietic (СD34, СD45) and epithelial (CD66е, CD326, рan keratin) cells increased of each passage. The data which obtained from immunophenotyping analysis culture myocardial cells were confirmed change morphology monolayer through fibroblast cells.

Conclusion and prospects for further research: 1. Primary culture of the rats myocardial cells heterogeneous and the dominant cell quantity epithelioid morphology. 2. During the cultivation cell culture changed epithelial-like cell morphology to fibroblast-like through a higher proliferation index of fibroblast-like cell. 3. Change of the expression of surface markers varies in each passage CD34, CD66e, CD326, pan-keratin increased significantly; SD95 and troponin I reduced significantly 4. During the cultivation we observed the emergence of expression of new surface markers (CD227 and pan-keratin), which indicate about the differentiation of cells culture and its acquisition of other properties and characteristics.

Keywords: cell culture, myocardium, immunophenotyping, culturing, morphology.

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