Three‐dimensional electrospun gelatin scaffold coseeded with embryonic stem cells and sertoli cells: A promising substrate for in vitro coculture system
2019
In this study, we present an electrospun gelatin (EG) scaffold to mimic the
extracellular matrix of the testis. The EG scaffold was synthesized by
electrospinning and crosslinked with glutaraldehyde vapor to decrease its
water solubility and degradation rate. The scanning electron microscope
micrographs showed the homogenous morphology of randomly aligned gelatin
fibers. The average diameter of gelatin fibers before and after crosslinking
was approximately 180 and 220 nm, respectively. Modulus, tensile strength,
and elongation at break values were as 161.8 ± 24.4 MPa, 4.21 ± 0.54 MPa, and
7.06 ± 2.12 MPa, respectively. The crosslinked EG showed 75.2% ± 4.5% weight
loss after 14 days with no changes in the pH value of degradation solution.
Cytobiocompatibility of the EG for sertoli cells and embryonic stem cells (ESCs)
was determined in vitro. Sertoli cells were isolated from mouse testis and
characterized by immunostaining and flow cytometry. The effects of EG on
proliferation and attachment of both sertoli cells and ESCs were examined. The
EG scaffolds exhibited no cytotoxicity for sertoli and ESCs. Both sertoli and
ESCs were well attached and grown on EG. Coculture of sertoli and ESCs on EG
showed better ESCs adhesion compared with ESCs alone. Our findings indicate
the potential of EG as a substrate for proliferation, adhesion, and coculture
of sertoli and ESCs and may be considered as a promising engineered
microenvironment for in vitro coculture system with the aim of guiding stem
cells differentiation toward sperm‐producing cells.
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