Gene Expression of Mouse Hippocampal Stem Cells Grown in a Galactose-Derived Molecular Gel Compared to In Vivo and Neurospheres

Abstract

Background: N-heptyl-D-galactonamide (GalC7) is a small synthetic carbohydrate derivative that forms a biocompatible supramolecular hydrogel. In this study, the objective was to analyze more in-depth how neural cells differentiate in contact with GalC7. Method: Direct (ex vivo) cells of the fresh hippocampus and culture (In vitro) of the primary cells were investigated. In vitro, investigation performed under three conditions: on culture in neurospheres for 19 days, on culture in GalC7 gel for 7 days, and on culture in both neurospheres and GalC7 gel. Total RNA was isolated with TRIzol from each group, Sox8, Sox9, Sox10, Dcx, and Neurod1 expression levels were measured by qPCR. Result: Sox8 and Sox10, oligodendrocyte markers, and Sox9, an astrocyte marker, were expressed at a much higher level after 7 days of culture in GalC7 hydrogel compared to all other conditions. Dcx, a marker of neurogenesis, and Neurod1, a marker of neuronal differentiation, were expressed at better levels in the GalC7 gel culture compared to the neurosphere. Conclusions: These results show that the GalC7 hydrogel brings different and interesting conditions for inducing the differentiation and maturation of neural progenitor cells compared with polymer-based scaffolds or cell-only conditions. The differences observed open new perspectives in tissue engineering, induction, and transcript analysis.

Dynamic Interplay Between miR-124-3p and EGF in Regulating The 'Giant' Phenotype by RNA Signalling Mechanism

“…The hippocampus was aspirated with an automatic pipette under a binocular microscope and taken into 1mL Hanks' Balanced Salt (HBS). After the hippocampal cells were homogenised with 0.25% Trypsin-EDTA (Gibco, Life Technologies), 24 well-plates coated with 300 mg Poly (2-hydroxyethyl methacrylate) (pHEMA) (Sigma, Germany) were placed on Dulbecco's Modi ed Eagle Medium: Nutrient Mixture F-12 (DMEM/F12) (Sigma, Germany) medium (Bayram et al, 2021). 20 ng/mL EGF and Insulin-Transferrin-Selenium-A (ITS) were added to one of the pairs.…”

“…Total RNA was isolated from the samples (E7 days old embryos, adult hippocampus, E19.5 days old hippocampus and NS that were cultured from hippocampal cells for both 12 1 in Supplementary 1). The data were analysed using the delta delta ct method and normalised using Beta-actin (9).…”

Wet spinning and 3D printing of supramolecular hydrogels in acid-base and dynamic conditions

“…and subtle changes of these triggers can strongly change the properties of the self-assembled objects and the resulting gels 16,[29][30][31][32][33][34][35][36][37][38] . Molecular hydrogels have been developed for various applications, including biomaterials 11,34,[39][40][41][42][43][44] , drug delivery 10 , 3D printing 45,46 , scaffolds for mineralization 47 , 48 etc. Because of the absence of covalent links between molecules, these hydrogels are quite fragile.…”

Wet spinning of a library of carbohydrate low molecular weight gels

“…This well-defined micro/nanostructuration is less often observed in polymer gels. More specifically when biocompatible molecular hydrogels are used as scaffolds for cell culture 10,11,12,13 what cells feel at the micro or nanometric scale will depend on the micro/nanostructuration, that is different from one gel to another. They can sense the stiffness but also the topography at the nano and microscale 14,15,16 , and even the chirality of the selfassembled fibers 17 .…”