IMMUNOPHENOTYPIC PROFILE OF STEM CELLS FROM THE BONE MARROWS OF RABBITS AT THE LATER PASSAGES IN VITRO
A.M.Maliuk
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National University of Life and Environmental Sciences of Ukraine, Heroiv Oborony str., 15; Kyiv, 03041, Ukraine
Despite the great interest in somatic stem cells from bone marrow of animals due to their multipotency, flexibility, simplicityof their obtaining and using, generally accepted methods for their isolation from tissues does not exist. In different laboratories use different methods of isolation and purification of stem cells, based on their size, adhesive properties, expression of surface antigens and inconnectionwiththisreasonthe receiving cell populations significantly differ in their biological characteristics.
Isolation and characterization of stem cells from bone marrow requires the development of specific marker antigens. A grate work has been done in this way, resulting in appearancesof many monoclonal antibodies. However, obtaining of a «pure» populations of multipotent stem cells due to a single marker remains a distant dream.
However, remains the question whether the stem cells of animal origin are heterogeneous at later passages during in vitro cultivation, because all studies related to CD-receptors system of stem cells conducted during their cultivation at early passages.
Multipotent stem cells for experimental studies were obtained from bone marrow of tubular bones of rabbit. Obtained cells wascultured in standard growthmedium. Microscopic analysis was carried out using inverted microscope Axiovert 40 (Carl Zeiss).
During immunocytochemical studies forthereason of examination the expression of specific markers, were used multipotentstem cellsfrom bone marrow of rabbit at seventh, twelfth and eighteenth passages. For studyof immunophenotype profile of stem cells were determined the expression of proteins associated with proliferation and cell cycle, apoptosis, cell adhesion and cytoskeleton.
Conducted immunocytochemical studies indicate that multipotent stem cells of the bone marrow of rabbits, which cultivates in standard growth media at later passages express markers of mesenchymal and epithelial cells, which confirms their heterogeneity and plasticity. At later passages dominates cells which express markers of mesenchymal origin was found.
Keywords: MULTIPOTENT STEM CELLS, BONE MARROW, MONOCLONAL ANTIBODIES IMMUNOCYTOCHEMICALANALYSIS, NUCLEAR PROTEINS, E-CADHERIN, N-CADHERIN, ACTIN, VIMENTIN
