IMPROVING METHODOLOGY OF ENDOTHELIAL CELL RESEARCH: SYNOPSIS AND PROSPECTS

Highlights The creation of a domestic bank of standardized endothelial cell cultures (arterial, venous, microvascular, valvular, and lymphatic lineages) and the development of an import-substituting technological process for working with them is a key task to enable next-generation endothelial physiology studies. The cultural and molecular justification for selecting the best source of autologous endothelial cells with the highest regenerative potential (in particular in context of their proliferative and angiogenic capacities) represents a primary task for endothelial physiology. The research methodology for studying endothelial cell physiology under normal and pathological conditions includes assessing their proliferative and angiogenic properties and molecular profiling. Abstract One of the primary goals in studying endothelial physiology is the development of a biomedical cell product in the form of autologous endothelial cells with high regenerative potential. These cells would be used to stimulate angiogenesis in ischemic tissues and to coat tissue-engineered constructs (such as vascular grafts) for their endothelialization prior to implantation into the human body. This review examines the key applied aspects of endothelial physiology aimed at achieving these objectives, including the selection of endothelial cell sources for modeling various pathological processes and regenerative medicine, the creation of a bank of standardized endothelial cell lines with various differentiation directions to enhance the effectiveness of this modeling, methods to ensure adhesion, sustained proliferation, and physiological functioning of primary endothelial cells in culture, as well as the methodology for assessing proliferative and angiogenic activity in the context of analyzing endothelial cell resistance to external influences. The discussion emphasizes the need for a comparative analysis of the regenerative properties of microvascular endothelial cells from subcutaneous adipose tissue, colony-forming endothelial cells from peripheral venous blood, and endothelial cells differentiated from induced pluripotent stem cells. Additionally, the current state of import substitution for various components of endothelial cell technology is reviewed, including standardized cell lines, functional protein coatings, and three-dimensional matrices for angiogenesis assessment, culture media for isolation, cultivation, and experimentation with endothelial cells, as well as other reagents for the isolation and subculturing of endothelial cells, culture dishes, and antibodies for flow cytometric and fluorescent microscopic immunophenotyping. The review also analyzes the prospects for import substitution of missing components, such as certain endothelial cell lines, magnetic beads with immobilized antibodies for their isolation, and antibodies conjugated with various fluorophores.