Three-dimensional mesenchymal stromal cells spheroids generated by RGD-induced self-assembly technique for tissue...

Area of Science:

• Biomaterials Science
• Cell Biology
• Tissue Engineering

Background:

• 3D spheroids from mesenchymal stromal cells (MSCs) offer in vivo-like conditions for tissue engineering.
• Current methods for spheroid generation can be complex.

Purpose of the Study:

• To develop a novel and simple approach for generating multicellular spheroids from MSCs.
• To utilize cell self-aggregation induced by a specific peptide for spheroid formation.

Main Methods:

• Mesenchymal stromal cells (MSCs) were cultured in monolayers.
• Cell self-aggregation was induced using the cyclo-RGDfK(TPP) peptide at concentrations of 5-100 µM.
• Spheroid formation was observed within 72 hours.
• Cell viability was confirmed using Calcein AM staining and confocal laser scanning microscopy.
• Reversibility of spheroids into single-cell cultures was assessed after peptide removal.

Main Results:

• The cyclo-RGDfK(TPP) peptide successfully induced MSC self-aggregation into multicellular spheroids within 72 hours.
• Vital dye staining confirmed high cell viability within the generated spheroids.
• The spheroids demonstrated reversibility, dissociating into single cells upon peptide removal.
• This reversibility facilitated cell invasion into collagen hydrogels.

Conclusions:

• A simple peptide-induced self-aggregation method effectively generates viable multicellular spheroids from MSCs.
• The ability to reverse spheroids into single cells is advantageous for subsequent tissue integration, such as hydrogel invasion.
• This spheroid-based cultivation approach holds significant potential for advancing tissue engineering strategies.