Abstract
Objective : to develop a model of a biomedical cell product that is consistent with the «homologous drug» strategy based on protocols for preparing the cell component and scaffold carrier for preclinical studies on a large laboratory animal (pig). Materials and methods . Biomedical cell products and skin equivalents (SE), were formed using plasma cryoprecipitate prepared from blood plasma of healthy donors and mesenchymal stem cells (MSCs) of human adipose tissue. Cryoprecipitate from pig blood plasma and human adipose tissue-derived MSCs were used to form model skin equivalents (mSE). Bright-field microscopy, phase-contrast microscopy (Leica DMI 3000B) and fluorescence microscopy (Cytation 5 imager; BioTek, USA) were used to monitor the state of cells in the culture and in the composition of the equivalents. Scaffolds for equivalents were tested for cytotoxicity (MTT test, direct contact method). The cell distribution density was characterized by author’s method (Patent No. 2675376 of the Russian Federation). Results . An mSE was developed for preclinical studies on a large laboratory animal (pig). In the mSE, components that change from halogen to xenogenic conditions during transplantation to the animal were replaced. A comprehensive approach to preparing mSE was presented. It includes sampling of primary pig biomaterial, extraction and characterization of adipose tissue-derived MSCs, preparation of a scaffold carrier for the corresponding «homologous drug» strategy. Cytotoxicity of the mSE scaffold was evaluated. It was shown that mSE provides mechanical support (similar to SE) to cells, as well as comparable development of cellular events during cultivation. Conclusion . A model of a biomedical cell product was developed. This model is consistent with the «homologous drug» strategy for preclinical studies on a large laboratory animal (pig). The paper presented a comprehensive approach to developing a model equivalent based on protocols for preparation and testing of the cellular component, the scaffold carrier and the ready-to-use model equivalent.
Highlights
Objective: to develop a model of a biomedical cell product that is consistent with the «homologous drug» strategy based on protocols for preparing the cell component and scaffold carrier for preclinical studies on a large laboratory animal
Biomedical cell products and skin equivalents (SE), were formed using plasma cryoprecipitate prepared from blood plasma of healthy donors and mesenchymal stem cells (MSCs) of human adipose tissue
An model skin equivalents (mSE) was developed for preclinical studies on a large laboratory animal
Summary
Все манипуляции по выделению и культивиро‐ ванию клеток, работы с кровью и ее производны‐ ми, работы по формированию и культивированию гидрогелевых скаффолдов проводили в помещении класса С в стерильных условиях ламинаров (класс А) в лаборатории биотехнологий ФГБОУ ВО «ПИМУ» Минздрава России. Включенный в исследование, пред‐ ставил добровольное информированное согласие
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