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

• Published in: Analytical and bioanalytical chemistry Vol. 402; no. 3; pp. 1083 - 1091
• Main Authors: Detwiler, David A., Dobes, Nicholas C., Sims, Christopher E., Kornegay, Joe N., Allbritton, Nancy L.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.01.2012; Springer
• Subjects: Acrylic acid; Analytical Chemistry; Animals; Arrays; Biochemistry; Bulging; Cell Adhesion; Cell Culture Techniques - instrumentation; Cell differentiation; Cell Proliferation; Cell Separation - instrumentation; Cells, Cultured; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Cloning; Collagen; Copolymers; Culture; Differentiation; Dogs; Epoxy resins; Food Science; Laboratory Medicine; Microstructure; Monitoring/Environmental Analysis; Muscle Cells - cytology; Optimization; Original Paper; Polystyrene; Polystyrenes - chemistry; Stem cells; Surface Properties; Tissue Array Analysis - instrumentation; Bioanalytical methods; Cell systems; Single cell analysis; Biomaterials
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-011-5596-9

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.