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Analytical and Bioanalytical Chemistry

, Volume 402, Issue 3, pp 1083–1091 | Cite as

Polystyrene-coated micropallets for culture and separation of primary muscle cells

  • David A. Detwiler
  • Nicholas C. Dobes
  • Christopher E. Sims
  • Joe N. Kornegay
  • Nancy L. AllbrittonEmail author
Original Paper

Abstract

Despite identification of a large number of adult stem cell types, current primary cell isolation and identification techniques yield heterogeneous samples, making detailed biological studies challenging. To identify subsets of isolated cells, technologies capable of simultaneous cell culture and cloning are necessary. Micropallet arrays, a new cloning platform for adherent cell types, hold great potential. However, the microstructures composing these arrays are fabricated from an epoxy photoresist 1002F, a growth surface unsuitable for many cell types. Optimization of the microstructures’ surface properties was conducted for the culture of satellite cells, primary muscle cells for which improved cell isolation techniques are desired. A variety of surface materials were screened for satellite cell adhesion and proliferation and compared to their optimal substrate, gelatin-coated Petri dishes. A 1-μm thick, polystyrene copolymer was applied to the microstructures by contact printing. A negatively charged copolymer of 5% acrylic acid in 95% styrene was found to be equivalent to the control Petri dishes for cell adhesion and proliferation. Cells cultured on control dishes and optimal copolymer-coated surfaces maintained an undifferentiated state and showed similar mRNA expression for two genes indicative of cell differentiation during a standard differentiation protocol. Experiments using additional contact-printed layers of extracellular matrix proteins collagen and gelatin showed no further improvements. This micropallet coating strategy is readily adaptable to optimize the array surface for other types of primary cells.

Keywords

Bioanalytical methods Biomaterials Cell systems Single cell analysis 

Notes

Acknowledgments

This research was supported by a grant from the National Institutes of Health (R01EB007612) to NLA, a Parent Project Muscular Dystrophy Weisman Fellowship to NCD, and an MDA infrastructure Grant to JNK. We are grateful to Pavak Shah for his microscopy assistance and Philip Gach for his guidance with manuscript preparation.

Supplementary material

216_2011_5596_MOESM1_ESM.pdf (77 kb)
Table S1 Optimized parameters for contact printing of various copolymers onto 1002F surfaces (PDF 76 kb)

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • David A. Detwiler
    • 1
  • Nicholas C. Dobes
    • 2
  • Christopher E. Sims
    • 2
  • Joe N. Kornegay
    • 1
  • Nancy L. Allbritton
    • 2
    • 3
    • 4
    Email author
  1. 1.Department of Pathology and Laboratory MedicineUniversity of North CarolinaChapel HillUSA
  2. 2.Department of ChemistryUniversity of North CarolinaChapel HillUSA
  3. 3.Department of Biomedical EngineeringUniversity of North CarolinaChapel HillUSA
  4. 4.Department of Biomedical EngineeringNorth Carolina State UniversityRaleighUSA

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