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<article article-type="research-article" dtd-version="1.3" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xml:lang="ru"><front><journal-meta><journal-id journal-id-type="publisher-id">regmedjournal</journal-id><journal-title-group><journal-title xml:lang="ru">Регенерация органов и тканей</journal-title><trans-title-group xml:lang="en"><trans-title>Регенерация органов и тканей</trans-title></trans-title-group></journal-title-group><issn pub-type="epub">2949-5938</issn><publisher><publisher-name>Общество регенеративной медицины</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.60043/2949-5938-2024-4-49-67</article-id><article-id custom-type="elpub" pub-id-type="custom">regmedjournal-90</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="ru"><subject>ОРИГИНАЛЬНЫЕ СТАТЬИ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>ORIGINAL ARTICLES</subject></subj-group></article-categories><title-group><article-title>Выделение и культивирование клеток эпителия эндометрия человека</article-title><trans-title-group xml:lang="en"><trans-title>Isolation and cultivation of human endometrial epithelial cells</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Кожухарова</surname><given-names>И. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Kozhukharova</surname><given-names>I. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Кожухарова Ирина Викторовна - к.б.н., старший научный сотрудник Лаборатории внутриклеточной сигнализации Отдела внутриклеточной сигнализации и транспорта</p><p>194064, Санкт-Петербург, Тихорецкий пр., 4</p></bio><bio xml:lang="en"><p>Irina V. Kozhukharova - Dr. Sci. (Biology), senior scientist of Laboratory of Intracellular Signaling, Department of Intracellular Signaling and Transport</p><p>St. Petersburg, Tikhoretsky ave., 4</p></bio><email xlink:type="simple">aldomnina@mail.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Кунеев</surname><given-names>И. К.</given-names></name><name name-style="western" xml:lang="en"><surname>Kuneev</surname><given-names>I. K.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Кунеев Иван Константинович - аспирант, младший научный сотрудник Лаборатории внутриклеточной сигнализации Отдела внутриклеточной сигнализации и транспорта</p><p>194064, Санкт-Петербург, Тихорецкий пр., 4</p></bio><bio xml:lang="en"><p>Ivan K. Kuneev - PhD student, junior scientist of Laboratory of Intracellular Signaling, Department of Intracellular Signaling and Transport</p><p>St. Petersburg, Tikhoretsky ave., 4</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Шорохова</surname><given-names>М. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Shorokhova</surname><given-names>M. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Шорохова Мария Александровна - к.б.н., научный сотрудник Лаборатории внутриклеточной сигнализации Отдела внутриклеточной сигнализации и транспорта</p><p>194064, Санкт-Петербург, Тихорецкий пр., 4</p></bio><bio xml:lang="en"><p>Maria A. Shorokhova - Dr. Sci. (Biology), scientist of Laboratory of Intracellular Signaling, Department of Intracellular Signaling and Transport</p><p>St. Petersburg, Tikhoretsky ave., 4</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Харченко</surname><given-names>М. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Kharchenko</surname><given-names>M. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Харченко Марианна Викторовна - к.б.н., старший научный сотрудник Лаборатории динамики внутриклеточных мембран Отдела внутриклеточной сигнализации и транспорта</p><p>194064, Санкт-Петербург, Тихорецкий пр., 4</p></bio><bio xml:lang="en"><p>Marianna V. Kharchenko - Dr. Sci. (Biology), senior scientist of Laboratory of Intracellular Membranes Dynamics, Department of Intracellular Signaling and Transport</p><p>St. Petersburg, Tikhoretsky ave., 4</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Корнилова</surname><given-names>Е. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Kornilova</surname><given-names>E. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Корнилова Елена Сергеевна - д.б.н., заведующий Лаборатории динамики внутриклеточных мембран Отдела внутриклеточной сигнализации и транспорта</p><p>194064, Санкт-Петербург, Тихорецкий пр., 4</p></bio><bio xml:lang="en"><p>Elena S. Kornilova - Dr. Sci. (Biology), head of Laboratory of Intracellular Membranes Dynamics, Department of Intracellular Signaling and Transport</p><p>St. Petersburg, Tikhoretsky ave., 4</p></bio><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Домнина</surname><given-names>А. П.</given-names></name><name name-style="western" xml:lang="en"><surname>Domnina</surname><given-names>A. P.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Домнина Алиса Павловна - к.б.н., старший научный сотрудник Лаборатории внутриклеточной сигнализации Отдела внутриклеточной сигнализации и транспорта</p><p>194064, Санкт-Петербург, Тихорецкий пр., 4</p></bio><bio xml:lang="en"><p>Alisa P. Domnina - Dr. Sci. ( Biology), senior scientist of L aboratory of Intracellular Signaling, Department of Intracellular Signaling and Transport</p><p>St. Petersburg, Tikhoretsky ave., 4</p></bio><xref ref-type="aff" rid="aff-1"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru"><institution>ФГБУН «Институт цитологии» Российской академии наук</institution><country>Россия</country></aff><aff xml:lang="en"><institution>Institute of Cytology of the Russian Academy of Sciences</institution><country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2024</year></pub-date><pub-date pub-type="epub"><day>19</day><month>12</month><year>2025</year></pub-date><volume>2</volume><issue>4</issue><fpage>49</fpage><lpage>67</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Кожухарова И.В., Кунеев И.К., Шорохова М.А., Харченко М.В., Корнилова Е.С., Домнина А.П., 2025</copyright-statement><copyright-year>2025</copyright-year><copyright-holder xml:lang="ru">Кожухарова И.В., Кунеев И.К., Шорохова М.А., Харченко М.В., Корнилова Е.С., Домнина А.П.</copyright-holder><copyright-holder xml:lang="en">Kozhukharova I.V., Kuneev I.K., Shorokhova M.A., Kharchenko M.V., Kornilova E.S., Domnina A.P.</copyright-holder><license xml:lang="ru" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>Данная работа распространяется под лицензией Creative Commons Attribution 4.0.</license-p></license><license xml:lang="en" license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>This work is licensed under a Creative Commons Attribution 4.0 License.</license-p></license></permissions><self-uri xlink:href="https://www.regmed-journal.ru/jour/article/view/90">https://www.regmed-journal.ru/jour/article/view/90</self-uri><abstract><p>Целью данной работы была разработка и оптимизация методов получения и культивирования клеток эпителия эндометрия человека из неинвазивного источника — менструальной крови. Исследование направлено на создание персонализированных моделей эндометрия in vitro для изучения механизмов имплантации эмбриона, поиска маркеров рецептивности эндометрия и разработки новых подходов к лечению гинекологических заболеваний, включая бесплодие.Методы. Для получения клеток эндометрия использовалась менструальная кровь, собранная от здоровых доноров. Фрагменты эндометрия выделялись с помощью фильтрации и ферментативной обработки. Культивирование клеток проводилось в различных условиях: в виде монослоя (2D), тканевой культуры и органоидов (3D). Для подтверждения эпителиального фенотипа использовалось иммуноцитохимическое окрашивание на маркеры цитокератин и Е-кадгерин. Для исследования эндоцитоза рецептора эпидермального фактора роста (EGFR) в тканевой культуре использовался флюоресцентно-меченный эпидермальный фактора роста (EGF).Результаты. Было показано, что менструальная кровь является доступным источником для получения жизнеспособных клеток эндометрия. Полученная тканевая культура эндометрия сохраняет архитектуру ткани и может служить моделью для исследования эндоцитоза рецептора эпидермального фактора роста (EGFR). Культивирование клеток эпителия эндометрия в виде органоидов позволило сохранить эпителиальный фенотип и пролиферативную активность клеток в течение длительного времени. Органоиды демонстрировали способность к самоорганизации и формированию однослойных клеточных структур, что подтверждает их пригодность для моделирования процессов, происходящих в эндометрии in vivo. В то же время культивирование в 2D-условиях приводило к быстрому старению клеток и потере их функциональных свойств.Заключение. Разработанные методы культивирования клеток эпителия эндометрия в виде тканевой культуры и органоидов открывают новые возможности для изучения механизмов имплантации эмбриона и поиска маркеров рецептивности эндометрия. Полученные результаты имеют большое значение для развития персонализированной медицины, в частности для повышения эффективности программ вспомогательных репродуктивных технологий (ВРТ) и разработки новых терапевтических подходов к лечению гинекологических заболеваний.</p></abstract><trans-abstract xml:lang="en"><p>The aim of this work was to develop and optimize methods for obtaining and culturing human endometrial epithelial cells from a non-invasive source — menstrual blood. The study aims to create personalized in vitro endometrial models to study embryo implantation mechanisms, search for markers of endometrial receptivity, and develop new approaches to treat gynecological diseases, including infertility.Methods. Menstrual blood collected from healthy donors was used to obtain endometrial cells. Endometrial fragments were isolated by filtration and enzymatic treatment. Cell culture was carried out under various conditions: as a monolayer (2D), tissue culture and organoids (3D). Immunocytochemical staining for the markers cytokeratin and E-cadherin was used to confirm the epithelial phenotype. To investigate endocytosis of the epidermal growth factor receptor ( EGFR) in tissue culture, fluorescence-labeled EGF was used.Results. Menstrual blood has been shown to be an affordable source for producing viable endometrial cells. The resulting endometrial tissue culture preserves tissue architecture and can serve as a model for the study of endocytosis of the epidermal growth factor receptor ( EGFR). Cultivation of endometrial epithelium cells in the form of organoids made it possible to preserve the epithelial phenotype and proliferative activity of cells for a long time. Organoids demonstrated the ability to self-organize and form single-layer cell structures, which confirms their suitability for modeling the processes occurring in the endometrium in vivo. At the same time, cultivation under 2D conditions led to rapid aging of cells and loss of their functional properties.Conclusion. The developed methods for culturing endometrial epithelial cells in the form of tissue culture and organoids open up new possibilities for studying the mechanisms of embryo implantation and searching for markers of endometrial receptivity. The findings have important implications for the development of personalized medicine, in particular for improving the effectiveness of assisted reproductive technology (ART) programs and developing new therapeutic approaches for the treatment of gynecological diseases.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>персонализированная медицина</kwd><kwd>эндометрий</kwd><kwd>эпителий эндометрия</kwd><kwd>органоиды</kwd><kwd>менструальная кровь</kwd><kwd>имплантация эмбриона</kwd><kwd>рецептивность эндометрия</kwd></kwd-group><kwd-group xml:lang="en"><kwd>personalized medicine</kwd><kwd>endometrium</kwd><kwd>endometrial epithelium</kwd><kwd>organoids</kwd><kwd>menstrual blood</kwd><kwd>embryo implantation</kwd><kwd>endometrial receptivity</kwd></kwd-group><funding-group><funding-statement xml:lang="ru">Исследование по выделению, культивированию и анализу клеток эпителия эндометрия человека выполнено за счет гранта Российского научного фонда № 22-74-10126. Исследование стимуляции эндоцитоза EGFR и выявление его локализации в клетках кокультуры эпителия и стромальных клеток эндометрия выпол- нено за счет гранта Российского научного фонда № 23-14-00335.</funding-statement><funding-statement xml:lang="en">The study on the isolation, cultivation and analysis of human endometrial epithelial cells was supported by Russian Science Foundation (grant No. 22-74-10126). The study of stimulation of endocytosis of EGFR and the identification of its localization in cells of co-culture of epithelium and stromal cells of the endometrium was supported by Russian Science Foundation (grant No. 23-14-00335).</funding-statement></funding-group></article-meta></front><back><ref-list><title>References</title><ref id="cit1"><label>1</label><citation-alternatives><mixed-citation xml:lang="ru">Goetz LH, Schork NJ. Personalized medicine: motivation, challenges, and progress. Fertil Steril. 2018 Jun;109(6):952–963. 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