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Generation of Haploid Spermatids on Silk Fibroin-Alginate-Laminin-Based Porous 3D Scaffolds Publisher Pubmed



Bashiri Z1, 2, 3 ; Moghaddaszadeh A4 ; Falak R5 ; Khadivi F6 ; Afzali A7 ; Abbasi M8 ; Sharifi AM1, 9 ; Asgari HR1, 2 ; Ghanbari F10 ; Koruji M1, 2
Authors
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Authors Affiliations
  1. 1. Stem cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  2. 2. Department of Anatomy, School of Medicine, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  3. 3. Omid Fertility & Infertility Clinic, Hamedan, 6516796198, Iran
  4. 4. Departement of Biomedical Engineering, Science and Research Branch, Islamic Azad University, Tehran, 1477893855, Iran
  5. 5. Immunology Research Center (IRC), Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  6. 6. Department of Anatomy, School of Medicine, Shahrekord University of Medical Sciences, Shahrekord, 8815713471, Iran
  7. 7. Hajar hospital, Shahrekord University of Medical Sciences, Shahrekord, 8816854633, Iran
  8. 8. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, 1417653761, Iran
  9. 9. Tissue Engineering Group (NOCERAL), Department of Orthopedics Surgery, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia
  10. 10. Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, 1449614535, Iran

Source: Macromolecular Bioscience Published:2023


Abstract

In vitro production of sperm is a desirable idea for fertility preservation in azoospermic men and prepubertal boys suffering from cancer. In this study, a biocompatible porous scaffold based on a triad mixture of silk fibroin (SF), alginate (Alg), and laminin (LM) is developed to facilitate the differentiation of mouse spermatogonia stem cells (SSCs). Following SF extraction, the content is analyzed by SDS-PAGE and stable porous 3D scaffolds are successfully prepared by merely Alg, SF, and a combination of Alg-SF, or Alg-SF-LM through freeze-drying. Then, the biomimetic scaffolds are characterized regarding the structural and biological properties, water absorption capacity, biocompatibility, biodegradability, and mechanical behavior. Neonatal mice testicular cells are seeded on three-dimensional scaffolds and their differentiation efficiency is evaluated using real-time PCR, flow cytometry, immunohistochemistry. Blend matrices showed uniform porous microstructures with interconnected networks, which maintained long-term stability and mechanical properties better than homogenous structures. Molecular analysis of the cells after 21 days of culture showed that the expression of differentiation-related proteins in cells that are developed in composite scaffolds is significantly higher than in other groups. The application of a composite system can lead to the differentiation of SSCs, paving the way for a novel infertility treatment landscape in the future. © 2023 Wiley-VCH GmbH.
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