Repository of Research and Investigative Information

Repository of Research and Investigative Information

Kurdistan University of Medical Sciences

Three-dimensional co-culture of human spermatogonial stem cells with Sertoli cells in soft agar culture system supplemented by growth factors and Laminin

(2020) Three-dimensional co-culture of human spermatogonial stem cells with Sertoli cells in soft agar culture system supplemented by growth factors and Laminin. Acta Histochemica.

Full text not available from this repository.

Official URL: https://www.scopus.com/inward/record.uri?eid=2-s2....

Abstract

Application of a three-dimensional (3D) culture system for in vitro proliferation and differentiation of human spermatogonial stem cells (SSCs) is a useful tool for the investigation of the spermatogenesis process and the management of male infertility particularly in prepubertal cancer patients. The main purpose of this study was to investigate the proliferation of human SSCs co-cultured with Sertoli cells in soft agar culture system (SACS) supplemented by Laminin and growth factors. Testicular cells were isolated from testes of brain-dead patients and cultured in two-dimensional (2D) culture system for 3 weeks. After 3 weeks, functional SSCs were evaluated by xenotransplantation and also identification of cells was assessed by immunocytochemistry, flow cytometry, and RT-PCR. Then, SSCs and Sertoli cells were transferred to the upper layer of SACS for 3 weeks. After 3 weeks, the number of colonies and the expression of specific SSCs and Sertoli cell markers, as well as apoptotic genes were evaluated. Our results showed that transplanted SSCs, migrated into the basement membrane of seminiferous tubules of recipient mice. The expression of PLZF, α6-Integrin, and Vimentin proteins in SSCs and Sertoli cells were observed in 2D and 3D culture systems. The expression rate of PLZF, α6-Integrin, Bcl2, and colony number in SACS supplemented by Laminin and growth factors group were significantly higher than non-supplemented groups (P ≤ 0.01), but the expression rate of c-kit and Bax in supplemented group were significantly lower than non-supplemented groups (P ≤ 0.05). This 3D co-culture system decreased apoptosis and increased propagation of human SSCs. Therefore, this designed system can be utilized to increase the proliferation of human SSCs in prepubertal male cancer and azoospermic men to obtain an adequate SSCs number to outotransplant success and in vitro spermatogenesis. © 2020

Item Type: Article
Keywords: agar; alpha6 integrin; cell marker; epidermal growth factor; fibroblast growth factor 2; glial cell line derived neurotrophic factor; growth factor; laminin; leukemia inhibitory factor; protein Bax; stem cell factor; vimentin; zinc finger and BTB domain containing protein 16, alpha 6 integrin gene; animal cell; animal experiment; animal tissue; apoptosis; Article; basement membrane; Bax gene; Bcl2 gene; brain death; c kit gene; cell count; cell culture; cell culture technique; cell function; cell isolation; cell migration; cell transplantation; coculture; controlled study; flow cytometry; functional assessment; genetic screening; human; human cell; human cell culture; human tissue; immunocytochemistry; male; mouse; nonhuman; PLZF gene; protein expression; real time polymerase chain reaction; seminiferous tubule; Sertoli cell; soft agar culture system; spermatogonium; stem cell; three dimensional coculture system; two dimensional culture system; xenotransplantation
Journal or Publication Title: Acta Histochemica
Volume: 122
Number: 5
Identification Number: 10.1016/j.acthis.2020.151572
Depositing User: مهندس جمال محمودپور
URI: http://eprints.muk.ac.ir/id/eprint/2292

Actions (login required)

View Item View Item