Search academic texts

Information × Registration Number 0224U033178, (0120U100381) , R & D reports Title Cryopreservation of spermatogonial stem and accessory regulatory cells of the testes with using of biopolymer gels. popup.stage_title Отримання та визначення морфофункціональних характеристик сперматогоніальних стовбурових та акцесорно-регуляторних клітин із кріоконсервованих фрагментів звитих канальців сім’яників Head Holtsev Anatolii M., Доктор медичних наук Registration Date 19-12-2024 Organization Institute of Cryobiology and Cryomedicine of the National Academy of Sciences of Ukraine popup.description1 Experimental reasoning of the possibility of using biopolymer gels to improve the preservation of the morphofunctional state of spermatogonial stem and accessory-regulatory cells of the testes of rat at the stages of cryopreservation popup.description2 The use of modern technologies of cryopreservation of testicular tissue samples in prepubertal patients is one of the ways to maintain their fertility in the future. The purpose of the study was to investigate the proliferative potential, morphological characteristics and expression of specific markers of cell culture obtained from cryopreserved and vitrified fragments of seminiferous tubules of rats’ testis. The isolation of cells from native, cryopreserved and vitrified fragments of seminiferous tubules of immature rats was performed by incubation in a solution of collagenase type IV (1 mg/mL) + DNase (500 μg/mL). Cell viability was determined by trypan blue staining. Monoclonal antibodies CD9-FITC, CD24-PE, CD45-FITC, CD90-FITC were used for immunophenotype analysis. Morphological characteristics, proliferative activity (MTT assay), relative number of cells positive for MAGE-B1 and vimentin were assessed in the obtained cultures. The analysis of phenotypic characteristics showed that cells from native, cryopreserved and vitrified fragments of seminiferous tubules were characterized by high expression level of CD9 (≥ 40 %), CD24 (≥ 70 %), CD90 (≥ 70 %) and low expression of the CD45 (≤ 1 %). In cell culture in vitro, the studied cells from cryopreserved and vitrified rat’s FSTs had the ability to adhere and proliferate while maintaining a cells population positive for MAGE-B1 and vimentin. The results can be the basis for the development of effective protocols for the cultivation and cryopreservation of testicular spermatogonial stem cells in order to restore fertility in men. Product Description popup.authors Volkova Nataliia O. Holtsev Anatolii M. Sokil Larysa V. Stepaniuk Liudmyla V. Chernyshenko Liudmyla H. Yukhta Mariia S. popup.nrat_date 2024-12-19 Close