Enzymatically crosslinked gelatin hydrogel promotes the proliferation of adipose tissue-derived stromal cells

Abstract

Gelatin hydrogel crosslinked by microbial transglutaminase (mTG) exhibits excellent performance in cell adhesion, proliferation, and differentiation. We examined the gelation time and gel strength of gelatin/mTG hydrogels in various proportions to investigate their physical properties and tested their degradation performances in vitro. Cell morphology and viability of adipose tissue-derived stromal cells (ADSCs) cultured on the 2D gel surface or in 3D hydrogel encapsulation were evaluated by immunofluorescence staining. Cell proliferation was tested via Alamar Blue assay. To investigate the hydrogel effect on cell differentiation, the cardiac-specific gene expression levelsof Nkx2.5, Myh6, Gja1, and Mef2c in encapsulated ADSCs with or without cardiac induction medium were detected by real-time RT-PCR. Cell release from the encapsulated status and cell migration in a 3D hydrogel model were assessed in vitro. Results show that the gelatin/mTG hydrogels are not cytotoxic and that their mechanical properties are adjustable. Hydrogel degradation is related to gel concentration and the resident cells. Cell growth morphology and proliferative capability in both 2D and 3D cultures were mainly affected by gel concentration. PCR result shows that hydrogel modulus together with induction medium affects the cardiac differentiation of ADSCs. The cell migration experiment and subcutaneous implantation show that the hydrogels are suitable for cell delivery.

Characterization of thin gelatin hydrogel membranes with balloon properties for dynamic tissue engineering

“…Previously, measurements of samples of 5% gelatin and comparable amounts of mTG as used here, have shown Young's modulus in the range of ~6 kPa measured on a Solid Analyzer at 37°C without having the sample immersed . Others have shown Young's modulus of 12.4 kPa of 5% gelatin hydrogels where the measurements were performed at room temperature . The lower modulus measured in the stretch setup (Figure ) compared to the rheometer and Solid Analyzer is believed to be caused by swelling of hydrogels in the immersed state.…”

“…This reduces time and cost and diminishes the technical difficulties in when casting stretchable hydrogel. Furthermore, mTG is non‐toxic, biocompatible, and is the most studied enzyme in protein‐based crosslinked hydrogels in tissue engineering . The mechanical behavior of gelatin crosslinking with mTG is dependent on the temperature at which the crosslinking is performed.…”

Microchannel network hydrogel induced ischemic blood perfusion connection

“…Since microbial transglutaminase (mTG) cross-links glutamine and lysine via a transamidation reaction, the mechanical properties of the gelatin hydrogel depended on the mTG dosage and the amounts of these molecules as presented by the gelatin concentration. Previous studies reported that the gelation time of mTG gel ranged from~15 s to hours with a constant mTG concentration (10 U g −1 ) [47][48][49] . The microchannel hydrogel induced invasion and M2 polarization of monocytes ( Fig.…”

Regulation of adipose‐tissue‐derived stromal cell orientation and motility in 2D‐ and 3D‐cultures by direct‐current electrical field

“…The isolation method of primary ADSCs was described in our previous publication (Li et al 2015;Yang et al 2016). Harvested ADSCs were seeded at a density of 1.0 9 10 4 cells/cm 2 on 0.1% gelatin-coated coverslips and placed into six-well tissue-culture plates (TCPs) and incubated at 37°C in a humid 5% CO 2 atmosphere for 24 h. Electrical stimulations were performed using homemade EF bioreactors, as described in our previous publication (Long et al 2011).…”