Tridimensional configurations of human mesenchymal stem/stromal cells to enhance cell paracrine potential towards wound healing processes

J Biotechnol. 2017 Nov 20;262:28-39. doi: 10.1016/j.jbiotec.2017.09.020. Epub 2017 Sep 28.


This study proposes to use alginate encapsulation as a strategy to assess the paracrine activity of 3D- and 2D-cultured human bone marrow mesenchymal stem/stromal cells (BM MSC) in the setting of wound repair and regeneration processes. A side-by-side comparison of MSC culture in three different 3D configurations (spheroids, encapsulated spheroids and encapsulated single cells) versus 2D monolayer cell culture is presented. The results reveal enhanced resistance to oxidative stress and paracrine potential of 3D spheroid-organized BM MSC. MSC spheroids (148±2μm diameter) encapsulated in alginate microbeads evidence increased angiogenic and chemotactic potential relatively to encapsulated single cells, as supported by higher secreted levels of angiogenic factors and by functional assays showing the capability of encapsulated MSC to promote formation of tubelike structures and migration of fibroblasts into a wounded area. In addition, a higher expression of the anti-inflammatory factor tumor necrosis factor alpha-induced protein 6 (TSG-6) was demonstrated by RT-PCR for encapsulated and non-encapsulated spheroids. Culture of spheroids within an alginate matrix maintains low aggregation levels below 5% and favors resistance to oxidative stress. These are important factors towards the establishment of more standardized and controlled systems, crucial to explore the paracrine effects of 3D-cultured MSC in therapeutic settings.

Keywords: Encapsulation; Mesenchymal stem/stromal cells (MSC); Paracrine activity; Spheroids; Wound healing.

MeSH terms

  • Alginates / chemistry*
  • Apoptosis
  • Cell Adhesion Molecules
  • Cell Culture Techniques*
  • Cell Line
  • Cell Survival
  • Cells, Cultured
  • Chemokine CXCL12 / metabolism
  • Cytokines / metabolism
  • Fibroblasts
  • Glucuronic Acid
  • Hepatocyte Growth Factor / metabolism
  • Hexuronic Acids
  • Humans
  • Interleukin-6
  • Mesenchymal Stem Cells / cytology*
  • Oxidative Stress
  • Spheroids, Cellular / cytology
  • Tumor Necrosis Factor-alpha
  • Vascular Endothelial Growth Factor A / metabolism
  • Wound Healing*


  • Alginates
  • CXCL12 protein, human
  • Cell Adhesion Molecules
  • Chemokine CXCL12
  • Cytokines
  • HGF protein, human
  • Hexuronic Acids
  • IL6 protein, human
  • Interleukin-6
  • TNF protein, human
  • TNFAIP6 protein, human
  • Tumor Necrosis Factor-alpha
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A
  • Hepatocyte Growth Factor
  • Glucuronic Acid