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Testis-derived macroporous 3D scaffold as a platform for the generation of mouse testicular organoids


The extracellular matrix-derived scaffolds provide an efficient platform for the generation of organ-like structures. Successful development of testicular organoids (TOs) with the capability of supporting complete spermatogenesis have not been reported yet. Here, we have developed an optimized method for decellularization of ram testicular tissue fragments. Our findings showed that testicular fragments treated with a serial combination of Triton X-100 and SDS in PBS for 48 h resulted in the efficient removal of cellular materials and retention of the extracellular matrix (ECM) components. In order to fabricate testis-derived scaffolds (TDSs), testicular ECM (T-ECM) was digested in acid/pepsin, followed by neutralization of pre-gel solution to form a hydrogel. Then, the hydrogels were freeze-dried and cross-linked using a chemical method. To reach the optimal concentration for T-ECM in the fabrication of TDS, the scaffold properties including porosity, pore size, swelling behavior, and degradation were evaluated. Our study suggested that 25 mg/ml of the T-ECM is the best concentration for the fabrication of macroporous TDS for demonstrating lower pore size, homogeneously distributed pores, and higher swelling ratio. Furthermore, inoculation of neonatal mouse testicular cells onto TDSs resulted in the generation of multicellular TOs in which the differentiation of spermatogonial cells to post-meiotic cells were confirmed. Hormonal analysis of TDS revealed the functionality of TOs in the secretion of testosterone and inhibin B. The current study also demonstrated macroporous TDS could provide a novel platform for testicular tissue engineering and in vitro spermatogenesis.

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Publication details

The article was received on 19 Aug 2018, accepted on 11 Jan 2019 and first published on 12 Jan 2019

Article type: Paper
DOI: 10.1039/C8BM01001C
Citation: Biomater. Sci., 2019, Accepted Manuscript
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    Testis-derived macroporous 3D scaffold as a platform for the generation of mouse testicular organoids

    T. Rezaei Topraggaleh, M. Rezazadeh Valojerdi, L. Montazeri and H. Baharvand, Biomater. Sci., 2019, Accepted Manuscript , DOI: 10.1039/C8BM01001C

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