3D-technology of the formation and maintenance of single dormant microspheres from 2000 human somatic cells and their reactivation in vitro

Bull Exp Biol Med. 2014 Nov;158(1):137-44. doi: 10.1007/s10517-014-2709-4. Epub 2014 Nov 19.

Abstract

We developed an original reproducible 3D-technology for preparation of single dormant microspheres consisting of 2000 somatic cells. The dynamics of microsphere assembly from mesenchymal and epithelial cells of retinal pigment epithelium was traced using time-lapse microscopy: formation of a loose aggregate over 24 h followed by its gradual consolidation and formation of a compact viable microsphere with a diameter of 100-150 μ by day 7. The cell number in the formed microspheres remains unchanged. Reactivation observed upon fusion of epithelial and/or mesenchymal microspheres results in the formation of a united compact microtissue. The fusion dynamics reproduces spherogenesis irrespective of the initial amount of co-cultured microspheres. Reactivation via two-step induced angiogenesis opens new prospects for production of vascularized microspheres and microtissues.

MeSH terms

  • Antigens, CD / metabolism
  • Cell Culture Techniques
  • Cell Differentiation
  • Epithelial-Mesenchymal Transition
  • Humans
  • Retinal Pigment Epithelium / cytology
  • Spheroids, Cellular / physiology*
  • Time-Lapse Imaging
  • Vascular Endothelial Growth Factor A / physiology

Substances

  • Antigens, CD
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A