Three-dimensional wet-electrospun poly(lactic acid)/multi-wall carbon nanotubes scaffold induces differentiation of human menstrual blood-derived stem cells into germ-like cells

Abstract

Infertility caused by the disruption or absence of germ cells is a major and largely incurable medical problem. Germ cells (i.e., sperm or egg) play a key role in the transmission of genetic and epigenetic information across generations. Generation of gametes derived in vitro from stem cells hold promising prospects which could potentially help infertile men and women. Menstrual blood-derived stem cells are a unique stem cell source. Evidence suggests that menstrual blood-derived stem cells exhibit a multi-lineage potential and have attracted extensive attention in regenerative medicine. To maintain the three-dimensional structure of natural extra cellular matrices in vitro, scaffolds can do this favor and mimic a microenvironment for cell proliferation and differentiation. According to previous studies, poly(lactic acid) and multi-wall carbon nanotubes have been introduced as novel and promising biomaterials for the proliferation and differentiation of stem cells. Some cell types have been successfully grown on a matrix containing carbon nanotubes in tissue engineering but there is no report for this material to support stem cells differentiation into germ cells lineage. This study designed a 3D wet-electrospun poly(lactic acid) and poly(lactic acid)/multi-wall carbon nanotubes composite scaffold to compare infiltration, proliferation, and differentiation potential of menstrual blood-derived stem cells toward germ cell lineage with 2D culture. Our primary data revealed that the fabricated scaffold has mechanical and biological suitable qualities for supporting and attachments of stem cells. The differentiated menstrual blood-derived stem cells tracking in scaffolds using scanning electron microscopy confirmed cell attachment, aggregation, and distribution on the porous scaffold. Based on the differentiation assay by RT-PCR analysis, stem cells and germ-like cells markers were expressed in 3D groups as well as 2D one. It seems that poly(lactic acid)/multi-wall carbon nanotubes scaffold-seeded menstrual blood-derived stem cells could be viewed as a novel, safe, and accessible construct for these cells, as they enhance germ-like generation from menstrual blood-derived stem cells.

Spermatogenesis induction of spermatogonial stem cells using nanofibrous poly(l‐lactic acid)/multi‐walled carbon nanotube scaffolds and naringenin

“…The FTIR spectra of PLLA/MWCNTs fibers (Figure A) showed the characteristic peaks of PLLA, related to the existence of C═O, C─O, ─CH 3 asymmetric, and ─CH 3 symmetric bonds. The absorbance peak at 1758 cm −1 is linked to C═O stretching bonds . Other stretching absorbance peaks in 1086, 2943, and 2944 cm −1 are related to the C─O, ─CH 3 asymmetric, and ─CH 3 symmetric bonds in PLLA nanofibers, respectively.…”

“…Besides, carbon‐based nanomaterials including graphene sheets, carbon nanofibers, carbon dots, and single/multi‐walled carbon nanotubes (CNTs) with great mechanical, electrical, and optical properties are vastly under investigation in the field of optimizing regenerative medicine scaffolds . In addition, CNTs can be incorporated into different biomaterials including medically approved polymers to construct highly conductive composite substrates . These nanoparticles are also great candidates for preparing the nanotopographic cues for determining the fate of stem cells.…”

Endometrial stem/progenitor cells and their roles in immunity, clinical application, and endometriosis

“…MenSCs partially (over 50%) express the pluripotency marker SSEA-4, but not Oct-4. MenSCs can differentiate into adipocytic [ 44 ], osteogenic [ 45 ], cardiomyocytic [ 46 ], and neurocytic lineages [ 47 ], as well as respiratory epithelial, endothelial, myocytic, hepatic [ 48 ], germ-like [ 49 , 50 ], and pancreatic cells [ 42 , 51 ] (Table 1 ). Replacement of fetal bovine serum with human platelet derivatives can promote the differentiation of MenSCs into osteoblasts [ 52 ].…”

“…MenSCs with properties of high survival rate in vivo and easy access make them very useful for stem cell transplantation in POF therapy. By two-dimensional culture and 3D scaffold culture system, MenSCs can differentiate into germ-like cells in vitro [ 49 , 50 ]. MenSCs transplantation increases the body weight of POF mice, improves the estrus cycle, and restores the fertility of POF mice [ 78 ].…”

“…The scaffold contains an extensive cartilage-like extracellular matrix whose glycosaminoglycan content is about 50% higher than that of the 2D culture system through which MenSCs differentiated [ 79 ]. On the 3D wet-electrospun poly (lactic acid)/multi-wall carbon nanotube scaffold, MenSCs can be differentiated into germ-like cells [ 50 ]. Based on the bilayer amniotic membrane/nano-fibrous fibroin scaffold, MenSCs can be differentiated into keratinocyte like cells in the presence of keratinocytes derived from human foreskin [ 80 ].…”