Russian Journal of PhysiologyRussian Journal of PhysiologyРоссийский физиологический журнал им. И.М. Сеченова0869-81392658-655XThe Russian Academy of Sciences67930810.31857/S0869813925020057UIWBQPEXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleEx vivo Murine Thromboinflammation Model with Validation on EMT-6 Breast CancerEx vivo модель тромбовоспаления у мышей с валидацией на модели рака молочной железы EMT-6KorobkinaJ. J. D.КоробкинаЮ. Д. Д.
Russian Federation
asve6nikova@yandex.ruMishukovA. A.МишуковА. А.
Russian Federation
asve6nikova@yandex.ruOsidakE. O.ОсидакЕ. О.
Russian Federation
asve6nikova@yandex.ruSveshnikovaA. N.СвешниковаА. Н.
Russian Federation
asve6nikova@yandex.ruCenter for Theoretical Problems of Physicochemical Pharmacology of the RASЦентр теоретических проблем физико-химической фармакологии РАНDmitry Rogachev National Research Center of Pediatric Hematology, Oncology and ImmunologyНациональный медицинский исследовательский центр детской гематологии, онкологии и иммунологии им. Дмитрия Рогачева16022025111226627709052025Copyright ©; 2025, Russian Academy of SciencesCopyright ©; 2025, Российская академия наук2025Russian Academy of SciencesРоссийская академия наукhttps://journals.eco-vector.com/0869-8139/article/view/679308

Thromboinflammation is a complex interaction between the hemostasis system and the immune system associated with the participation of neutrophils in the process of thrombosis. An imbalance in the mutual activation of platelets and neutrophils in various pathologies leads to thrombosis or bleeding. Previously, we developed a technique for ex vivo observation of the process of thrombosis and chemotaxis of neutrophils in parallel-plate flow chambers coated with fibrillar collagen. The aim of this work was to develop a technique for ex vivo observation of the thromboinflammation process in mice, which allows analyzing the interaction of polymorphonuclear leukocytes (PMN) with growing blood clots. To validate the technique, a breast cancer model was used in BALB/c mice (7 days after orthotopic inoculation with EMT-6 tumor cell culture). In the blood samples of healthy mice, blood clots formed on fibrillar collagen, however, nuclear cells were not observed. The use of a combination of fibronectin and collagen as a substrate made it possible to induce thrombosis and monitor the movement and behavior of PMNs in the vicinity of blood clots. Using the developed model, it was shown that in breast cancer, the growth of blood clots is slowed down – the relative size of blood clots in breast cancer is 21 ± 11% of the field of view compared with 39 ± 10% in healthy mice. At the same time, the number of PMNs adhering to blood clots and the speed of their movement on the substrate do not differ in healthy mice and mice with tumors. However, the number of PMNs leaving the thrombus and sliding onto the fibronectin-collagen matrix was significantly increased in mice with a tumor (39 ± 23 versus 15 ± 8 in healthy controls). Thus, using the developed thromboinflammation model, it is shown that already at the early stages of tumor development, violations of the thromboinflammation process are observed.

Тромбовоспаление – это комплексное взаимодействие между системой гемостаза и иммунной системой, связанное с участием нейтрофилов в процессе тромбообразования. Дисбаланс взаимной активации тромбоцитов и нейтрофилов при различных патологиях приводит к тромбозам или кровотечениям. Ранее нами была разработана методика ex vivo наблюдения процесса тромбообразования и хемотаксиса нейтрофилов в плоскопараллельных проточных камерах, покрытых фибриллярным коллагеном. Целью данной работы была разработка методики ex vivo наблюдения процесса тромбовоспаления у мышей, позволяющая анализировать взаимодействие полиморфоядерных лейкоцитов (ПМЯЛ) с растущими тромбами. Для валидации методики использовалась модель рака молочной железы у мышей линии BALB/с (7 дней после ортотопической инокуляции культурой опухолевых клеток EMT-6). В образцах крови здоровых мышей на фибриллярным коллагене образовывались полноценные тромбы, однако ядерные клетки не наблюдались. Использование комбинации фибронектина и коллагена в качестве подложки позволило индуцировать тромбообразование и отслеживать движение и поведение ПМЯЛ в окрестности тромбов. При помощи разработанной модели показано, что при онкологии рост тромбов замедлен – относительный размер тромбов при раке молочной железы составляет 21 ± 11% от поля зрения по сравнению с 39 ± 10 % у здоровых мышей. При этом количество ПМЯЛ, адгезирующих к тромбам, и скорость их движения на подложке не отличаются у здоровых мышей и мышей с опухолями. Однако количество ПМЯЛ, покидающих тромб и сползающих на фибронектин-коллагеновый матрикс, значимо увеличено у мышей с опухолью (39 ± 23 против 15 ± 8 у здоровых контролей). Таким образом, с помощью разработанной модели тромбовоспаления показано, что уже на ранних этапах развития опухоли наблюдаются нарушения процесса тромбовоспаления.

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