kpcczКомплексные проблемы сердечно-сосудистых заболеванийComplex Issues of Cardiovascular Diseases2306-12782587-9537Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”10.17802/2306-1278-2024-13-3-118-129kpccz-1523Research ArticleАНАЛИТИЧЕСКИЙ ОБЗОР. Патологическая физиологияREVIEWS. Pathological physiologyАКТУАЛЬНЫЕ ПРОБЛЕМЫ МЕТОДОЛОГИИ ИЗУЧЕНИЯ НОРМАЛЬНОЙ И ПАТОЛОГИЧЕСКОЙ ФИЗИОЛОГИИ ЭНДОТЕЛИАЛЬНЫХ КЛЕТОК В КУЛЬТУРЕIMPROVING METHODOLOGY OF ENDOTHELIAL CELL RESEARCH: SYNOPSIS AND PROSPECTShttps://orcid.org/0000-0002-1518-3888ШишковаДарья КирилловнаShishkovaDaria K.

кандидат биологических наук заведующая лабораторией молекулярной, трансляционной и цифровой медицины отдела экспериментальной медицины федерального государственного бюджетного научного учреждения «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний» Кемерово, Российская Федерация

PhD, Head of the Laboratory for Molecular, Translational, and Digital Medicine, Department of Experimental Medicine, Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”, Kemerovo, Russian Federation

shishkovadk@gmail.comhttps://orcid.org/0000-0002-1746-8895ФроловАлексей ВитальевичFrolovAlexey V.

доктор медицинских наук старший научный сотрудник лаборатории рентгенэндоваскулярной и реконструктивной хирургии сердца и сосудов отдела хирургии сердца и сосудов федерального государственного бюджетного научного учреждения «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний» Кемерово, Российская Федерация

MD, PhD, Senior Researcher, Laboratory for Endovascular and Reconstructive Cardiovascular Surgery, Department of Cardiovascular Surgery, Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”, Kemerovo, Russian Federation

kjerne@yandex.ruhttps://orcid.org/0000-0002-6652-5745МарковаВиктория ЕвгеньевнаMarkovaVictoria E.

младший научный сотрудник лаборатории молекулярной, трансляционной и цифровой медицины отдела экспериментальной медицины федерального государственного бюджетного научного учреждения «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний» Кемерово, Российская Федерация

Junior Researcher, Laboratory for Molecular, Translational, and Digital Medicine, Department of Experimental Medicine, Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”, Kemerovo, Russian Federation

marvika97@gmail.comhttps://orcid.org/0009-0007-6734-3787МарковаЮлия ОлеговнаMarkovaYulia O.

младший научный сотрудник лаборатории молекулярной, трансляционной и цифровой медицины отдела экспериментальной медицины федерального государственного бюджетного научного учреждения «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний» Кемерово, Российская Федерация

Junior Researcher, Laboratory for Molecular, Translational, and Digital Medicine, Department of Experimental Medicine, Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”, Kemerovo, Russian Federation

yulikmimi19@gmail.comhttps://orcid.org/0000-0002-1867-6354ЛазебнаяАнастасия ИвановнаLazebnayaAnastasia I.

младший научный сотрудник лаборатории молекулярной, трансляционной и цифровой медицины отдела экспериментальной медицины федерального государственного бюджетного научного учреждения «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний» Кемерово, Российская Федерация

Junior Researcher, Laboratory for Molecular, Translational, and Digital Medicine, Department of Experimental Medicine, Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”, Kemerovo, Russian Federation

https://orcid.org/0000-0001-8679-4857КутихинАнтон ГеннадьевичKutikhinAnton G.

доктор медицинских наук заведующий отделом экспериментальной медицины федерального государственного бюджетного научного учреждения «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний» Кемерово, Российская Федерация

MD, PhD, Head of the Department of Experimental Medicine, Federal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”, Kemerovo, Russian Federation

antonkutikhin@gmail.comФедеральное государственное бюджетное научное учреждение «Научно-исследовательский институт комплексных проблем сердечно-сосудистых заболеваний»РоссияFederal State Budgetary Institution “Research Institute for Complex Issues of Cardiovascular Diseases”Russian Federation202430092024133118129Copyright © Шишкова Д.К., Фролов А.В., Маркова В.Е., Маркова Ю.О., Лазебная А.И., Кутихин А.Г., 20242024Шишкова Д.К., Фролов А.В., Маркова В.Е., Маркова Ю.О., Лазебная А.И., Кутихин А.Г.Shishkova D.K., Frolov A.V., Markova V.E., Markova Y.O., Lazebnaya A.I., Kutikhin A.G.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.nii-kpssz.com/jour/article/view/1523Основные положения

Основные положения

 

 

Резюме

Резюме

Одной из основных целей изучения физиологии эндотелия является разработка биомедицинского клеточного продукта в виде аутологичных эндотелиальных клеток с высоким регенеративным потенциалом для стимуляции ангиогенеза в ишемизированных тканях, а также для покрытия ими тканеинженерных конструкций (к примеру, сосудистых протезов) для их эндотелизации перед имплантацией в организм человека. В данном обзоре рассматриваются основные прикладные аспекты физиологии эндотелия, направленные на достижение данных целей, а именно: выбор источника эндотелиальных клеток для моделирования различных патологических процессов и регенеративной медицины, создание банка стандартизированных линий эндотелиальных клеток с различными направлениями дифференцировки для повышения эффективности данного моделирования, способы обеспечения адгезии, устойчивой пролиферации и физиологического функционирования первичных эндотелиальных клеток в культуре, а также методология оценки пролиферативной и ангиогенной активности в контексте анализа устойчивости эндотелиальных клеток к внешним воздействиям. В процессе обсуждения обосновывается необходимость сравнительного анализа регенеративных свойств микрососудистых эндотелиальных клеток из подкожной жировой ткани, колониеформирующих эндотелиальных клеток из периферической венозной крови и эндотелиальных клеток, дифференцированных из индуцированных плюрипотентных стволовых клеток. Отдельно рассматривается текущее состояние процесса импортозамещения различных компонентов технологий работы с эндотелиальными клетками (стандартизированных клеточных линий, функционального белкового покрытия и трехмерного матрикса для оценки ангиогенеза, питательных сред для выделения, культивирования и проведения экспериментов с эндотелиальными клетками, иных реагентов для выделения и субкультивирования эндотелиальных клеток, культуральной посуды, антител для проточно-цитометрического и флюоресцентно-микроскопического иммунофенотипирования). Анализируются перспективы импортозамещения недостающих звеньев работы: ряда линий эндотелиальных клеток, магнитных бус с иммобилизованными антителами для их выделения, конъюгированных с различными флюорофорами антител.

Highlights

Highlights

 

 

Abstract

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.

Эндотелиальные клеткиПролиферацияАнгиогенезРегенеративная медицинаПитательные средыИммунофенотипированиеИмпортозамещениеEndothelial cellsProliferationAngiogenesisRegenerative medicineCell culture mediumImmunophenotypingImport substitution industrializationРабота выполнена при поддержке комплексной программы фундаментальных научных исследований СО РАН в рамках фундаментальной темы НИИ КПССЗ № 0419-2024-0001 «Разработка новых фармакологических подходов к экспериментальной терапии атеросклероза, технологий серийного производства реактивов и расходных материалов для изучения физиологии и патофизиологии сердечно-сосудистой системы и программного обеспечения на основе искусственного интеллекта для автоматизированной диагностики патологий системы кровообращения и автоматизированного расчета сердечно-сосудистого риска» и финансовой поддержке Министерства науки и высшего образования Российской Федерации в рамках национального проекта «Наука и университеты».ReferencesCrampton S.P., Davis J., Hughes C.C. 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The authors declare that there are no conflicts of interest present.