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Influence of discrete and continuous culture conditions on human mesenchymal stem cell lineage choice in RGD concentration gradient hydrogels.

Publication ,  Journal Article
Smith Callahan, LA; Policastro, GM; Bernard, SL; Childers, EP; Boettcher, R; Becker, ML
Published in: Biomacromolecules
September 2013

Stem cells have shown lineage-specific differentiation when cultured on substrates possessing signaling groups derived from the native tissue. A distinct determinant in this process is the concentration of the signaling motif. While several groups have been working actively to determine the specific factors, concentrations, and mechanisms governing the differentiation process, many have been turning to combinatorial and gradient approaches in attempts to optimize the multiple chemical and physical parameters needed for the next advance. However, there has not been a direct comparison between the cellular behavior and differentiation of human mesenchymal stem cells cultured in gradient and discrete substrates, which quantitates the effect of differences caused by cell-produced, soluble factors due to design differences between the culture systems. In this study, the differentiation of human mesenchymal stem cells in continuous and discrete polyethylene glycol dimethacrylate (PEGDM) hydrogels containing an RGD concentration gradient from 0 to 14 mM were examined to study the effects of the different culture conditions on stem-cell behavior. Culture condition was found to affect every osteogenic (alkaline phosphatase, Runx 2, type 1 collagen, bone sailoprotein, and calcium content) and adipogenic marker (oil red and peroxisome proliferator-activated receptor gamma) examined regardless of RGD concentration. Only in the continuous gradient culture did RGD concentration affect human mesenchymal stem-cell lineage commitment with low RGD concentrations expressing higher osteogenic differentiation than high RGD concentrations. Conversely, high RGD concentrations expressed higher adipogenic differentiation than low RGD concentrations. Cytoskeletal actin organization was only affected by culture condition at low RGD concentrations, indicating that it played a limited role in the differences in lineage commitment observed. Therefore, the role of discrete versus gradient strategies in high-throughput experimentation needs to be considered when designing experiments as we show that the respective strategies alter cellular outcomes even though base scaffolds have similar material and chemical properties.

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Published In

Biomacromolecules

DOI

EISSN

1526-4602

ISSN

1525-7797

Publication Date

September 2013

Volume

14

Issue

9

Start / End Page

3047 / 3054

Related Subject Headings

  • Polymers
  • Osteogenesis
  • Oligopeptides
  • Mesenchymal Stem Cells
  • Hydrogels
  • Humans
  • Extracellular Matrix Proteins
  • Cell Nucleus
  • Cell Culture Techniques
  • Antigens, Differentiation
 

Citation

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Smith Callahan, L. A., Policastro, G. M., Bernard, S. L., Childers, E. P., Boettcher, R., & Becker, M. L. (2013). Influence of discrete and continuous culture conditions on human mesenchymal stem cell lineage choice in RGD concentration gradient hydrogels. Biomacromolecules, 14(9), 3047–3054. https://doi.org/10.1021/bm4006112
Smith Callahan, Laura A., Gina M. Policastro, Sharon L. Bernard, Erin P. Childers, Ronna Boettcher, and Matthew L. Becker. “Influence of discrete and continuous culture conditions on human mesenchymal stem cell lineage choice in RGD concentration gradient hydrogels.Biomacromolecules 14, no. 9 (September 2013): 3047–54. https://doi.org/10.1021/bm4006112.
Smith Callahan LA, Policastro GM, Bernard SL, Childers EP, Boettcher R, Becker ML. Influence of discrete and continuous culture conditions on human mesenchymal stem cell lineage choice in RGD concentration gradient hydrogels. Biomacromolecules. 2013 Sep;14(9):3047–54.
Smith Callahan, Laura A., et al. “Influence of discrete and continuous culture conditions on human mesenchymal stem cell lineage choice in RGD concentration gradient hydrogels.Biomacromolecules, vol. 14, no. 9, Sept. 2013, pp. 3047–54. Epmc, doi:10.1021/bm4006112.
Smith Callahan LA, Policastro GM, Bernard SL, Childers EP, Boettcher R, Becker ML. Influence of discrete and continuous culture conditions on human mesenchymal stem cell lineage choice in RGD concentration gradient hydrogels. Biomacromolecules. 2013 Sep;14(9):3047–3054.
Journal cover image

Published In

Biomacromolecules

DOI

EISSN

1526-4602

ISSN

1525-7797

Publication Date

September 2013

Volume

14

Issue

9

Start / End Page

3047 / 3054

Related Subject Headings

  • Polymers
  • Osteogenesis
  • Oligopeptides
  • Mesenchymal Stem Cells
  • Hydrogels
  • Humans
  • Extracellular Matrix Proteins
  • Cell Nucleus
  • Cell Culture Techniques
  • Antigens, Differentiation