The Study of the Interaction of Mesenchymal Stem Cells and the Tumor Using the Methods of Fluorescent Bioimaging
The aim of the investigation is to study the pathogenesis of tumor development when interacting with mesenchymal stem cells transfected by a gene of red fluorescent protein using the method of vital bioimaging.
Materials and Methods. There were used adipose-derived adult stem (ADAS) cells taken from human adipose tissue. ADAS were transfected by a gene of red fluorescent protein Turbo FP635 (Close Joint Stock Company “Eurogene”, Russia) by the method of lentiviral transfection. Tumors were implanted to nude mice by subcutaneous injection of Hela Kyoto tumor cells (cervical cancer). ADAS labeled by fluorescent protein were injected to animals at different stages of carcinogenesis (0 and 8 days after tumor culture injection) by various ways: locally — in the tumor node forming region, and systemically — intravenously, in the tail vein. There were formed the following groups of animals: 1st group — an early stage of carcinogenesis (immediately after the injection) and systemic injection of ADAS; 2nd group — an early stage of carcinogenesis and local injection of ADAS; 3rd group — the stage of developed tumor (8 days) and systemic injection of ADAS. The control group consisted of the animals with induced tumor without stem cells injection.
Results. ADAS isolated and characterized with the help of immunocytochemical analysis had the phenotype of mesenchymal stem cells (there were expressed CD105, CD49d, STRO-1) and were differentiated in vitro in adipogenic, osteogenic, and chondrogenic approaches in induction media cultivation. The efficiency of transfection of ADAS by red fluorescent protein Turbo FP635 was 75%. The stem cells under study — ADAS labeled by red fluorescent protein Turbo FP635 in systemic injection were shown to be able to migrate in spleen, and in systemic and local injection — in bone marrow, lungs, and recipient’s tumor tissues. The methods of fluorescent bioimaging and laser scanning microscopy can be used to study the interaction between the tumor and mesenchymal stem cells. They effectively complement each other in gaining general knowledge of the distribution of migratory fluorescent cells.
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