Heparin stabilizes FGF-2 and modulates striatal precursor cell behavior in response to EGF

Exp Neurol. 2004 Aug;188(2):408-20. doi: 10.1016/j.expneurol.2004.05.007.

Abstract

Fibroblast and epidermal growth factors (FGF-2 and EGF) are powerful mitogens for neural precursor cells isolated from the developing striatum and grown as neurospheres. However, questions remain as to the exact role of each of these molecules, and how the proteoglycan heparin may modify their behavior. Here, we show that FGF-2 is remarkably unstable in culture media, but that heparin could completely prevent its degradation, which led to faster cell growth rates. In addition, heparin significantly increased the number of cells within the E14 striatum responding to a brief pulse of FGF-2. In contrast, EGF was unable to stimulate the growth of E14 striatal precursors. However, EGF could induce the division of E18 striatal precursors as neurospheres and acted synergistically with FGF-2. FGF-2/heparin neurospheres generated significantly more neurons than EGF neurospheres. Interestingly, the addition of heparin to EGF neurospheres, which had no effects on EGF stability or growth rates, increased the numbers of neurons generated to that seen for FGF-2/heparin neurospheres. EGF neurospheres were found to produce FGF-2, but addition of heparin did not affect its concentration within cells or in the medium suggesting this released FGF-2 may already be bound to a proteoglycan. In addition, expanding cells with EGF plus heparin in the presence of an FGF-2 blocker did not have a significant effect on the number of neurons generated confirming that the increase in neuronal number is through a mechanism which is independent of FGF-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Count
  • Cell Division / drug effects
  • Cells, Cultured
  • Clone Cells / cytology
  • Clone Cells / drug effects
  • Epidermal Growth Factor / pharmacology*
  • Fibroblast Growth Factor 2 / drug effects*
  • Fibroblast Growth Factor 2 / metabolism
  • Gestational Age
  • Heparin / metabolism
  • Heparin / pharmacology
  • Heparin / physiology*
  • Neostriatum / cytology
  • Neostriatum / drug effects*
  • Neostriatum / embryology
  • Rats
  • Rats, Sprague-Dawley
  • Spheroids, Cellular / cytology
  • Spheroids, Cellular / drug effects
  • Stem Cells / cytology
  • Stem Cells / drug effects*
  • Temperature

Substances

  • Fibroblast Growth Factor 2
  • Epidermal Growth Factor
  • Heparin