Isolation and Characteristics of Colony-Forming Endothelial Cells From Peripheral Blood in Patients With Ischemic Heart Disease

Матвеева, В. Г.; Khanova, Mariam Yu.; Великанова, Е. А.; Антонова, Л. В.; Сардин, Е. С.; Krutitsky, S. S.; Barbarash, O. L.

doi:10.31116/tsitol.2018.08.03

Cited by 2 publications

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“…После полимеризации трубчатый каркас погружали в фосфатно-солевой буфер-ный раствор с ε-аминокапроновой кислотой в объеме 2 мг/мл до последующего заселения клетками. Для заселения использовали культуру колониеформирующих эндотелиальных клеток (КФЭК), которую получали культивированием мононуклеарной фракции из крови пациентов с ишемической болезнью сердца в питательной среде EGM-2MV (Lonza, Швеция) с содержанием 5% эмбриональной бычьей сыворотки [3]. На внутреннюю поверхность биодеградируемых протезов с фибриновым покрытием вносили 7 × 10 5 КФЭК и культивировали в течение двух суток в статических условиях в СО 2 -инкубаторе при 37 °С.…”

Section: материалы и методыunclassified

Development of personalized cell-populated vascular graft in vitro

Khanova

Великанова

Глушкова

et al. 2021

Kompleks. probl. serdečno-sosud. zabol.

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Aim. To create a personalized cell-populated small-diameter vascular prosthesis in a pulsating bioreactor.Methods. Tubular grafts were made by electrospinning from mixtures of biodegradable polymers, poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(εcaprolactone) (PCL). The inner surface is modified with fibrin. Tubular scaffolds were colonized with cultured colony-forming endothelial cells and grown under static conditions for 2 days. Then, the cell-populated prostheses continued to be cultivated for 5 days in a pulsating bioreactor system with a final shear stress of 2.85 dynes/cm².Results. The advantages of the cultivation of cell-populated vascular prostheses in a pulsating bioreactor have been revealed. The selected mode of cultivation of cellpopulated vascular prostheses under conditions of a pulsating flow with a shear stress of 2.85 dynes/cm² did not have a damaging effect on the integrity of the endothelial monolayer. Moving unidirectional mechanical stimuli of chaotic orientation fibers of F-actin changed to a predominant orientation in the direction of flow, and also increased the expression of F-actin, Talin focal adhesion protein, and specific endothelial markers CD309, CD31, vWF.Conclusion. The creation of a personalized cell-populated small-diameter vascular prosthesis with a functional endothelial monolayer is possible due to the use of autologous endothelial cells, autologous fibrin, and cultivation under conditions of a pulsating flow.

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Section: материалы и методыunclassified

Development of personalized cell-populated vascular graft in vitro

Khanova

Великанова

Глушкова

et al. 2021

Kompleks. probl. serdečno-sosud. zabol.

Self Cite

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Influence of different concentrations of fibrinogen on the properties of a fibrin matrix for vascular tissue engineering

Матвеева

Геннадьевна²,

Khanova

et al. 2021

I.P. Pavlov Russian Medical Biological Herald

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Aim. To evaluate the potential utility of fibrin matrices containing 10, 20, and 25 mg/ml of fibrinogen (fibrin-10, fibrin-20, and fibrin-30, respectively) in vascular tissue engineering (VTE). Materials and Methods. Fibrinogen was isolated using the method of ethanol cryoprecipitation and polymerized using a solution of thrombin and CaCl2. The fibrin structure was studied in a scanning electron microscope, and the physical and mechanical properties of the material were tested on a Zwick/Roell test machine. The metabolic activity of endothelial cells (EC) on the fibrin surface was evaluated by the MTT assay, and the viability of fibroblasts in the thickness of fibrin and possibility for migration by in fluorescent and light microscopy. Percent of fibrin shrinkage was determined from the difference in the sample volumes before and after removal of moisture. Results. The fiber diameter did not differ among all fibrin samples, but the pore diameter in fibrin-30 was smaller than those in fibrin-10 and fibrin-20. A possibility for migration of fibroblasts into the depth of the fibrin matrix and preservation of 97-100% viability of cells at a depth 5 mm was confirmed. The metabolic activity of EC on the surface of fibrin-20 and fibrin-30 exceeded that on collagen, fibronectin, and fibrin-10. All fibrin samples shrank in volume to 95.5-99.5%, and the highest shrinkage was seen in fibrin-10. The physical and mechanical properties of fibrin were inferior to those of human A. mammaria by a factor of 10. Conclusion. Fibrin with fibrinogen concentrations of 20 and 30 mg/ml maintains a high metabolic and proliferative activity of EC on the surface and also a high viability of fibroblasts in the matrix. Its availability, ease of preparation, and a number of other favorable properties make fibrin a promising material for VTE. However, the problem of insufficient strength requires further investigations.

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Isolation and Characteristics of Colony-Forming Endothelial Cells From Peripheral Blood in Patients With Ischemic Heart Disease

Cited by 2 publications

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Development of personalized cell-populated vascular graft in vitro

Development of personalized cell-populated vascular graft in vitro

Influence of different concentrations of fibrinogen on the properties of a fibrin matrix for vascular tissue engineering

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