Patterning the differentiation of C2C12 skeletal myoblasts

Piyush Bajaj, Bobby Reddy, Larry Millet, Chunan Wei, Pinar Zorlutuna, Gang Bao, Rashid Bashir

Research output: Contribution to journalArticle

Abstract

Mammalian cells are sensitive to the physical properties of their micro-environment such as the stiffness and geometry of the substrate. It is known that the stiffness of the substrate plays a key role in the process of mammalian myogenesis. However, the effect of geometrical constraints on the process of myogenic differentiation needs to be explored further. Here, we show that the geometrical cues of substrates can significantly influence the differentiation process of C2C12 skeletal myoblasts. Three different geometries including lines of different widths, tori of different inner diameters, and hybrid structures (linear and circular features with different arc degrees) were created by micro-contact printing of fibronectin on the surface of Petri dishes. The differentiation of C2C12 cells was studied over a period of seven days and was quantified; we report the differentiation parameters of (1) fusion index, (2) degree of maturation, (3) alignment, and (4) response to electrical pulse stimulation (EPS). Hybrid structures with the smallest arc degree (hybrid 30°) showed the best results for all four differentiation parameters. The hybrid 30° pattern exhibits an ∼2-fold increase in the fusion index when compared to the line patterns and an ∼3-fold increase when compared to the toroid patterns. The hybrid 30° also showed a higher maturation index compared to the line or the toroid patterns. In response to electrical stimulation (20 V, 50 ms pulse, 1 Hz), mature myotubes on hybrid 30° patterns showed an ∼2-fold increase in cellular displacement when compared to myotubes on the line and torus patterns. We tested the influence of C2C12 cell density on fusion and maturation indices, and the results suggest that density does not exert significant influence on cellular differentiation under these conditions. Our results can have important implications in engineering skeletal muscle tissues and designing muscle cell bio-actuators.

Original languageEnglish (US)
Pages (from-to)897-909
Number of pages13
JournalIntegrative Biology
Volume3
Issue number9
DOIs
StatePublished - Sep 1 2011

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Skeletal Myoblasts
Fusion reactions
Muscle
Substrates
Cells
Stiffness
Geometry
Skeletal Muscle Fibers
Fibronectins
Electric Stimulation
Printing
Actuators
Physical properties
Tissue
Muscle Development
Muscle Cells
Cues
Cell Differentiation
Skeletal Muscle
Cell Count

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

Cite this

Bajaj, P., Reddy, B., Millet, L., Wei, C., Zorlutuna, P., Bao, G., & Bashir, R. (2011). Patterning the differentiation of C2C12 skeletal myoblasts. Integrative Biology, 3(9), 897-909. https://doi.org/10.1039/c1ib00058f

Patterning the differentiation of C2C12 skeletal myoblasts. / Bajaj, Piyush; Reddy, Bobby; Millet, Larry; Wei, Chunan; Zorlutuna, Pinar; Bao, Gang; Bashir, Rashid.

In: Integrative Biology, Vol. 3, No. 9, 01.09.2011, p. 897-909.

Research output: Contribution to journalArticle

Bajaj, P, Reddy, B, Millet, L, Wei, C, Zorlutuna, P, Bao, G & Bashir, R 2011, 'Patterning the differentiation of C2C12 skeletal myoblasts', Integrative Biology, vol. 3, no. 9, pp. 897-909. https://doi.org/10.1039/c1ib00058f
Bajaj P, Reddy B, Millet L, Wei C, Zorlutuna P, Bao G et al. Patterning the differentiation of C2C12 skeletal myoblasts. Integrative Biology. 2011 Sep 1;3(9):897-909. https://doi.org/10.1039/c1ib00058f
Bajaj, Piyush ; Reddy, Bobby ; Millet, Larry ; Wei, Chunan ; Zorlutuna, Pinar ; Bao, Gang ; Bashir, Rashid. / Patterning the differentiation of C2C12 skeletal myoblasts. In: Integrative Biology. 2011 ; Vol. 3, No. 9. pp. 897-909.
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