title: Engineering spatial control of multiple differentiation fates within a stem cell population.
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.
authors.affiliation: Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA 15213, USA.
authors.forename: Elmer D F
authors.initials: ED
authors.lastname: Ker
authors.forename: Bur
authors.initials: B
authors.lastname: Chu
authors.forename: Julie A
authors.initials: JA
authors.lastname: Phillippi
authors.forename: Burhan
authors.initials: B
authors.lastname: Gharaibeh
authors.forename: Johnny
authors.initials: J
authors.lastname: Huard
authors.forename: Lee E
authors.initials: LE
authors.lastname: Weiss
authors.forename: Phil G
authors.initials: PG
authors.lastname: Campbell
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D002478:Cells, Cultured; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects; D051379:Mice; D032446:Myoblasts / Q000166:cytology / Q000187:drug effects; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D016133:Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects*; D013710:Tendons / Q000166:cytology
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D016222:Fibroblast Growth Factor 2