Sustained levels of FGF2 maintain undifferentiated stem cell cultures with biweekly feeding

PLoS One. 2013;8(2):e56289. doi: 10.1371/journal.pone.0056289. Epub 2013 Feb 20.

Abstract

An essential aspect of stem cell culture is the successful maintenance of the undifferentiated state. Many types of stem cells are FGF2 dependent, and pluripotent stem cells are maintained by replacing FGF2-containing media daily, while tissue-specific stem cells are typically fed every 3rd day. Frequent feeding, however, results in significant variation in growth factor levels due to FGF2 instability, which limits effective maintenance due to spontaneous differentiation. We report that stabilization of FGF2 levels using controlled release PLGA microspheres improves expression of stem cell markers, increases stem cell numbers and decreases spontaneous differentiation. The controlled release FGF2 additive reduces the frequency of media changes needed to maintain stem cell cultures, so that human embryonic stem cells and induced pluripotent stem cells can be maintained successfully with biweekly feedings.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects*
  • Cells, Cultured
  • Cells, Immobilized / cytology
  • Cells, Immobilized / drug effects
  • Culture Media / pharmacology
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects
  • Embryonic Stem Cells / enzymology
  • Enzyme Activation / drug effects
  • Fibroblast Growth Factor 2 / pharmacology*
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / enzymology
  • Lactic Acid
  • MAP Kinase Signaling System / drug effects
  • Mice
  • Microspheres
  • Mitogen-Activated Protein Kinases / metabolism
  • Neural Stem Cells / cytology
  • Neural Stem Cells / drug effects
  • Neural Stem Cells / enzymology
  • Polyglycolic Acid
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Stem Cells / enzymology

Substances

  • Culture Media
  • Fibroblast Growth Factor 2
  • Polylactic Acid-Polyglycolic Acid Copolymer
  • Polyglycolic Acid
  • Lactic Acid
  • Mitogen-Activated Protein Kinases

Grant support

This work was supported by funding from the Regenerative Research Foundation, Rensselaer NY. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.