Engineering spatial control of multiple differentiation fates within a stem cell population

Biomaterials. 2011 May;32(13):3413-22. doi: 10.1016/j.biomaterials.2011.01.036. Epub 2011 Feb 12.

Abstract

The capability to engineer microenvironmental cues to direct a stem cell population toward multiple fates, simultaneously, in spatially defined regions is important for understanding the maintenance and repair of multi-tissue units. We have previously developed an inkjet-based bioprinter to create patterns of solid-phase growth factors (GFs) immobilized to an extracellular matrix (ECM) substrate, and applied this approach to drive muscle-derived stem cells toward osteoblasts 'on-pattern' and myocytes 'off-pattern' simultaneously. Here this technology is extended to spatially control osteoblast, tenocyte and myocyte differentiation simultaneously. Utilizing immunofluorescence staining to identify tendon-promoting GFs, fibroblast growth factor-2 (FGF-2) was shown to upregulate the tendon marker Scleraxis (Scx) in C3H10T1/2 mesenchymal fibroblasts, C2C12 myoblasts and primary muscle-derived stem cells, while downregulating the myofibroblast marker α-smooth muscle actin (α-SMA). Quantitative PCR studies indicated that FGF-2 may direct stem cells toward a tendon fate via the Ets family members of transcription factors such as pea3 and erm. Neighboring patterns of FGF-2 and bone morphogenetic protein-2 (BMP-2) printed onto a single fibrin-coated coverslip upregulated Scx and the osteoblast marker ALP, respectively, while non-printed regions showed spontaneous myotube differentiation. This work illustrates spatial control of multi-phenotype differentiation and may have potential in the regeneration of multi-tissue units.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / drug effects*
  • Cell Line
  • Cells, Cultured
  • Fibroblast Growth Factor 2 / pharmacology
  • Fluorescent Antibody Technique
  • Humans
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Mice
  • Myoblasts / cytology
  • Myoblasts / drug effects
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Polymerase Chain Reaction
  • Stem Cells / cytology*
  • Stem Cells / drug effects*
  • Tendons / cytology

Substances

  • Intercellular Signaling Peptides and Proteins
  • Fibroblast Growth Factor 2