Embryonic stem cells differentiated into neuron‐like cells using SB431542 small molecule on nanofibrous PLA/CS/Wax scaffold

Abstract

Electrospun nanofibrous scaffolds show huge potential to improve the neurological outcome in central nervous system disorders. In this study, we cultured mouse embryonic stem cells (mESCs) on an electrospun nanofibrous polylactic acid/ Chitosan/Wax (PLA/CS/Wax) scaffold and surveyed the attachment, behavior, and differentiation of mESCs into neural cells. Differentiation in neural-like cells (NLCs) was investigated with a medium containing SB431542 as a small molecule and conjugated linolenic acid after 20 days. We used Immunocytochemistry and quantitative real-time polymerase chain reaction (RT-PCR) techniques to assess neural marker expression in differentiated cells. SEM imaging demonstrated that mESCs could strongly attach, stretch, and differentiate on PLA/CS/Wax scaffolds. MESCs that were cultured on PLA/CS/Wax scaffolds showed enhanced numbers of neural structures and neural markers including Nestin, NF-H, Tuj-1, and Map2 in neural induction medium compared to the control sample. These results revealed that electrospun PLA/CS/Wax scaffolds associated with the induction medium can assemble proper conditions for stem cell differentiation into NLCs. We hope that the development of new technologies in neural tissue engineering may pave a new avenue for neural tissue regeneration. K E Y W O R D S beeswax, electrospun nanofibrous scaffolds, mESCs, neural differentiation, small molecule

Producing Human Amniotic Epithelial Cells-only Membrane for Transplantation

“…Generally, mammalian cells have a high demand of a proper support surface. Thus, some material such as gelatin, collagen matrix and matrigel (BD, USA) were used as support surface for culture of pluripotent stem cells like embryonic stem cells or induced pluripotent stem cells[49-52]. The BM of hAM was mainly consisted of collagen and proven to be a good supporting scaffold [12,53].…”

Electrospun Nanofiber Scaffold for Skin Tissue Engineering: A Review

“…67 The hydrophilic nature of PLA-CS fibers was found to have increased, thereby promoting the proliferation and osteogenic differentiation of marrow mesenchymal stem cells on the fibers. 69,70 The combination of PLGA with multiwalled CNTs and BC was found to enhance osteogenic differentiation and facilitate regeneration. 71 The study revealed that incorporating soluble eggshell membrane protein led to enhanced spinnability and mechanical strength of PLGA.…”

“…Materials made from synthetic polymers exhibit high spinnability and favorable mechanical properties. , Nevertheless, they are unable to enhance cell adhesion, proliferation, and differentiation, whereas natural polymers typically exhibit superior bioactivities despite having inferior mechanical strength. , Researchers have shown a tendency to combine synthetic and natural polymers using cospinning, coaxial electrospinning, and surface coating modification. , CS, GEL, and COL are incorporated to markedly decrease the hydrophobic nature of nanofibers and enhance cell adhesion . The hydrophilic nature of PLA-CS fibers was found to have increased, thereby promoting the proliferation and osteogenic differentiation of marrow mesenchymal stem cells on the fibers. , The combination of PLGA with multiwalled CNTs and BC was found to enhance osteogenic differentiation and facilitate regeneration . The study revealed that incorporating soluble eggshell membrane protein led to enhanced spinnability and mechanical strength of PLGA …”

Physical Cue‐Based Strategies on Peripheral Nerve Regeneration

“…At the cellular level, neural stem cells cultured on well‐arranged PLA fibers showed improved protuberances growth. [ 224 ] Similarly, SCs cultured on aligned PCL fibers, were observed to have their cell cytoskeleton and nuclei aligned and elongated along the fiber axes and, in addition, have the propensity of promoting SCs maturation. [ 225,226 ] As an explanation for the above phenomenon, differential gene expression of neurotrophic factors were observed in aligned, random, and smooth films.…”