Optimizing culture medium composition to improve oligodendrocyte progenitor cell yields in vitro from subventricular zone-derived neural progenitor cell neurospheres

PLoS One. 2015 Apr 2;10(4):e0121774. doi: 10.1371/journal.pone.0121774. eCollection 2015.

Abstract

Neural Stem and Progenitor Cells (NSC/NPC) are gathering tangible recognition for their uses in cell therapy and cell replacement therapies for human disease, as well as a model system to continue research on overall neural developmental processes in vitro. The Subventricular Zone is one of the largest NSC/NPC niches in the developing mammalian Central Nervous System, and persists through to adulthood. Oligodendrocyte progenitor cell (OPC) enriched cultures are usefull tools for in vitro studies as well as for cell replacement therapies for treating demyelination diseases. We used Subventricular Zone-derived NSC/NPC primary cultures from newborn mice and compared the effects of different growth factor combinations on cell proliferation and OPC yield. The Platelet Derived Growth Factor-AA and BB homodimers had a positive and significant impact on OPC generation. Furthermore, heparin addition to the culture media contributed to further increase overall culture yields. The OPC generated by this protocol were able to mature into Myelin Basic Protein-expressing cells and to interact with neurons in an in vitro co-culture system. As a whole, we describe an optimized in vitro method for increasing OPC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Becaplermin
  • Biomarkers / metabolism
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Coculture Techniques
  • Culture Media / chemistry
  • Culture Media / pharmacology*
  • Gene Expression
  • Genes, Reporter
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Heparin / pharmacology
  • Humans
  • Lateral Ventricles / cytology
  • Lateral Ventricles / drug effects*
  • Lateral Ventricles / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Myelin Basic Protein / genetics
  • Myelin Basic Protein / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects*
  • Neural Stem Cells / metabolism
  • Neurons / cytology
  • Neurons / drug effects*
  • Neurons / metabolism
  • Oligodendroglia / cytology
  • Oligodendroglia / drug effects*
  • Oligodendroglia / metabolism
  • Platelet-Derived Growth Factor / pharmacology
  • Primary Cell Culture
  • Proto-Oncogene Proteins c-sis / pharmacology

Substances

  • Biomarkers
  • Culture Media
  • Myelin Basic Protein
  • Platelet-Derived Growth Factor
  • Proto-Oncogene Proteins c-sis
  • enhanced green fluorescent protein
  • platelet-derived growth factor A
  • Green Fluorescent Proteins
  • Becaplermin
  • Heparin

Grants and funding

This work was supported by PIP 1767—Consejo Nacional de Investigaciones Científicas y Técnicas (http://www.conicet.gov.ar/) (PGF); PIP 0711—Consejo Nacional de Investigaciones Científicas y Técnicas (http://www.conicet.gov.ar/) (PGF); Préstamo BID PICT 2011-2040—Agencia Nacionasl de Promoción Científica y Tecnológica (http://www.agencia.mincyt.gob.ar/) (PGF); and Préstamo BID PICT 2008-0791)- Agencia Nacionasl de Promoción Científica y Tecnológica (http://www.agencia.mincyt.gob.ar/) (JMP). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.