Three dimensional neuronal cell cultures more accurately model voltage gated calcium channel functionality in freshly dissected nerve tissue

PLoS One. 2012;7(9):e45074. doi: 10.1371/journal.pone.0045074. Epub 2012 Sep 25.

Abstract

It has been demonstrated that neuronal cells cultured on traditional flat surfaces may exhibit exaggerated voltage gated calcium channel (VGCC) functionality. To gain a better understanding of this phenomenon, primary neuronal cells harvested from mice superior cervical ganglion (SCG) were cultured on two dimensional (2D) flat surfaces and in three dimensional (3D) synthetic poly-L-lactic acid (PLLA) and polystyrene (PS) polymer scaffolds. These 2D- and 3D-cultured cells were compared to cells in freshly dissected SCG tissues, with respect to intracellular calcium increase in response to high K(+) depolarization. The calcium increases were identical for 3D-cultured and freshly dissected, but significantly higher for 2D-cultured cells. This finding established the physiological relevance of 3D-cultured cells. To shed light on the mechanism behind the exaggerated 2D-cultured cells' functionality, transcriptase expression and related membrane protein distributions (caveolin-1) were obtained. Our results support the view that exaggerated VGCC functionality from 2D cultured SCG cells is possibly due to differences in membrane architecture, characterized by uniquely organized caveolar lipid rafts. The practical implication of use of 3D-cultured cells in preclinical drug discovery studies is that such platforms would be more effective in eliminating false positive hits and as such improve the overall yield from screening campaigns.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Calcium Channels / metabolism*
  • Caveolae / physiology
  • Caveolin 1 / genetics
  • Caveolin 1 / metabolism
  • Cell Culture Techniques*
  • Coated Materials, Biocompatible / chemistry
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism
  • Gene Expression
  • Ion Channel Gating / physiology
  • Lactic Acid / chemistry
  • Mice
  • Molecular Imaging
  • Polyesters
  • Polymers / chemistry
  • Polystyrenes / chemistry
  • Superior Cervical Ganglion / cytology
  • Superior Cervical Ganglion / metabolism*
  • Tissue Scaffolds

Substances

  • Calcium Channels
  • Caveolin 1
  • Coated Materials, Biocompatible
  • Polyesters
  • Polymers
  • Polystyrenes
  • Lactic Acid
  • poly(lactide)
  • DNA-Directed RNA Polymerases
  • Calcium

Grant support

This work was partly supported by National Science Foundation (0304340) and UGA Engineering Grants. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding received for this study.