Generation of Haploid Spermatids on Silk Fibroin-Alginate-Laminin-Based Porous ۳D Scaffolds

Background: In vitro production of sperm is one of the mos timportant options for fertility preservation in azoospermic menand prepubertal boys with cancer. Therefore, in this s tudy, abiocompatible porous scaffold based on silk fibroin-Alg containinglaminin was developed to differentiate mouse spermatogonias tem cells (SSCs).Materials and Methods: After extraction and characterizationof silk fibroin using SDS-PAGE analysis, s table porous ۳Dscaffolds were successfully prepared from combined solutionsthrough a freeze-dried method. Then, s tructural and biologicalproperties, biocompatibility, water absorption, degradabilityand mechanical behavior of biomimic scaffolds were characterized.Neonatal mice tes ticular cells were seeded on ۳D scaffoldsafter confirmation of nature and their differentiation efficiencywas evaluated using Real Time‐PCR, flow cytometry, immunohistochemis try techniques and H & E s taining. The function ofLeydig and Sertoli cells was also assessed using ELISA.Results: Blend matrices showed uniform porous micros tructurewith interconnected network, which significantly maintainedlong-term weight and better mechanical properties thanpure s tructures. The results of molecular analysis after ۲۱ daysof culture showed that the expression of differential markers(Acrosin, Scp۳ and Prm۱) in the ۳D sys tem containing lamininwas significantly higher than other groups. Hormonal analysisconfirmed the function of Leydig and Sertoli cells for the synthesisof tes tos terone and inhibin.Conclusion: The usage of a ۳D sys tem containing laminincould lead to the differentiation of SSCs and the progression ofmeiosis to the s tage of haploid spermatid that pave the way fornew human infertility treatments in the future.

کلیدواژه ها:

Spermatogonia S tem Cells ، Porous Scaffold ، Laminin