6-O-Sulfated Chitosan Promoting the Neural Differentiation of Mouse Embryonic Stem Cells

Abstract

Embryonic stem cells (ESCs) can be induced to differentiate into nerve cells, endowing them with potential applications in the treatment of neurological diseases and neural repair. In this work, we report for the first time that sulfated chitosan can promote the neural differentiation of ESCs. As a type of sulfated glycosaminoglycan analog, sulfated chitosan with well-defined sulfation sites and a controlled degree of sulfation (DS) were prepared through simple procedures and the influence of sulfated glycosaminoglycan on neural differentiation of ESCs was investigated. Compared with other sulfation sites, 6-O-sulfated chitosan showed the most optimal effects. By monitoring the expression level of neural differentiation markers using immunofluorescence staining and PCR, it was found that neural differentiation was better enhanced by increasing the DS of 6-O-sulfated chitosan. However, increasing the DS by introducing another sulfation site in addition to the 6-O site to chitosan did not promote neural differentiation as much as 6-O-sulfated chitosan, indicating that compared with DS, the sulfation site is more important. Additionally, the optimal concentration and incubation time of 6-O-sulfated chitosan were investigated. Together, our results indicate that the sulfate site and the molecular structure in a sulfated polysaccharide are very important for inducing the differentiation of ESCs. Our findings may help to highlight the role of sulfated polysaccharide in inducing the neural differentiation of ESCs.

Recent Advances in Artificially Sulfated Polysaccharides for Applications in Cell Growth and Differentiation, Drug Delivery, and Tissue Engineering

“…2.2.1. Applications in cell growth and differentiation : As a mimetic to overcome the limitations of heparin and heparan sulfate, CHS exhibits intrinsic biological properties towards cell differentiation, proliferation, and growth, according to the advances reported within the last decade …”

“…In 2014, Ding et al. reported, for the first time, that CHS could promote the neural differentiation of embryonic stem cells (ESCs; Figure ) . Compared with other sulfation sites, 6‐O‐sulfated chitosan showed the most optimal effects.…”

“…Applications in cell growth and differentiation:A samimetic to overcome the limitations of heparin and heparan sulfate, [10][11][12][13] CHS exhibitsi ntrinsic biological properties towards cell differentiation,p roliferation, andg rowth, according to the advances reported within the last decade. [79][80][81][82][83][84][85][86][87][88][89] In 2010, Ho et al reported thiol-modified CHS nanoparticles for the protection and release of FGF-2. [80] L929 fibroblast culture testss howed that thiol-modified 2-N-and 3,6-O-sulfated, 6-O-carboxymethyl chitosan/chitosan (N,O-SOCC/CHS) nanoparticles could effectively protect FGF-2 from inactivation over a1 20 hp eriod.…”

Selective Modification of Chitin and Chitosan: En Route to Tailored Oligosaccharides

“…Moreover, they have been applied in different areas, such as metal ion adsorption or drug delivery systems . Thus, the chemo‐ and regioselective control of sulfation is required to access compounds with specific characteristics . Unfortunately, and despite the wide number of articles covering chitin/chitosan sulfation, there is still a lack of reports with a comprehensive study on selective sulfation.…”

“…Chitosans sulfated at O‐6 are relevant biologic targets, and their preparation in high selectivity is of great importance . Chitosans sulfated at O‐6 have been obtained by using chlorosulfonic acid under acidic conditions .…”