Suspension-adapted Chinese hamster ovary-derived cells expressing green fluorescent protein as a screening tool for biomaterials

Abstract

Synthetic biomaterials play an important role in regenerative medicine. To be effective they must support cell attachment and proliferation in addition to being non-toxic and non-immunogenic. We used a suspension-adapted Chinese hamster ovary-derived cell line expressing green fluorescent protein (GFP) to assess cell attachment and growth on synthetic biomaterials by direct measurement of GFP-specific fluorescence. To simplify operations, all cell cultivation steps were performed in orbitallyshaken, disposable containers. Comparative studies between this GFP assay and previously established cell quantification assays demonstrated that this novel approach is suitable for rapid screening of a large number of samples. Furthermore the utility of our assay system was confirmed by evaluation of cell growth on three polyvinylidene fluoride polymer scaffolds that differed in pore diameter and drawing conditions. The data presented here prove the general utility of GFP-expressing cell lines and orbital shaking technology for the screening of biomaterials for tissue engineering applications.

Biocatalysis, Enzyme Engineering and Biotechnology

“…Mammalian cells are the ideal candidate for expression hosts (Engelhardt et al 2009, Hacker et al 2009) when posttranslational modifications (N-and O-glycosylation, disulfide bond formation) are a critical factor for the efficacy of the expressed protein (Baldi et al 2007, Werner et al 2007, Durocher and Butler 2009, Geisse and Fux 2009, Hacker et al 2009. Despite substantial limitations, such as high cost, low yield, instability of expression and time-consuming, a significant number of proteins (e.g.…”