291589 Cell Adhesion As a Function of LCST, Comparing Pnipam to A Novel Thermoresponsive Substrate

Monday, October 29, 2012
Hall B (Convention Center )
Ashley Kaminski, Tyler Fruneaux, Kevin Ling and Lauren S. Anderson, Chemical and Biomolecular Engineering, Lafayette College, Easton, PA

Cell adhesion to novel materials is determined by material surface characteristics, protein adsorption, and the cellular phenotypic response, all of which are interrelated.  Thermoresponsive polymer brushes offer a unique culture system whereby cells are released by a simple temperature switch.  PNIPAM, the gold standard for thermoresponsive polymers exhibits a lower critical solution temperature (LCST) of 32°C.  At temperatures below the LCST, polymer chains are soluble and expand; above the LCST, chains are insoluble and collapse.  A novel thermoresponsive substrate prepared from random copolymers of P(MEO2MA-co-OEGMA) (PMO)  has a distinct advantage over PNIPAM: the ability to tune the LCST to values between 26 and 90°C by varying the co-monomer ratio.  Changes in cell morphology and phenotype were assessed for both thermoresponsive systems compared to TCP and as a function of LCST.  Mouse fibroblasts were cultured on brushes for 48 hours and then harvested for gene expression analysis using real time RT-PCR.  Cell circularity and spreading were evaluated as a function of time to gain understanding of attachment efficiency. After 48 hours, cell circularity was statistically similar in all substrates and spreading was 90%+ despite significant differences at earlier time points.  Gene expression results suggest that phenotype is a function of LCST.

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