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

Roman Akasov1, Oxana Selina1, Tatyana Tolstova2

  • 1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Rus Acad Sci, Miklukho-Maklaya Str. 16/10, Moscow, 117997 Russia.

Cytotechnology
|April 9, 2026
PubMed
Summary

Researchers developed a simple method using a peptide to create multicellular spheroids from mesenchymal stromal cells (MSCs). This technique shows promise for tissue engineering applications.

Keywords:
Cyclo-RGD peptideMesenchymal stromal cellsMulticellular spheroidsScaffoldTissue engineering

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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.