A potential cell affinity sorbent: Fibronectin carrying poly(EGDMA/HEMA) microbeads

Abstract

Both non-swellable and swellable poly(EGDMA/HEMA) microbeads were produced by suspension copolymerization. These microbeads were modified by immobilization of a spacer-arm (hexamethylene diamine, HMDA) and fibronectin. The optimal values for modifications were as follows: the sodium periodate concentration 1.0 mg ml-1; the HMDA concentration 4 mg ml-1; and the glutaraldehyde concentration 0.070 μg ml-1. Adsorption of fibronectin onto the plain and periodate-oxidized poly(EGDMA/HEMA) microbeads were very similar, and were 0.025-0.035 mg fibronectin per g polymer, respectively. Fibronectin immobilization on poly(EGDMA/HEMA) microbeads were studied at different temperature, time and pH using single protein solution containing different amount of proteins. The optimal values for immobilizations were as follows: the initial fibronectin concentration 0.1 mg ml; temperature + 25°C; pH 7; the immobilization time 120 min. Both fibroblastic 3T3 and epithelial MDBK cells were attached to these unmodified and modified microbeads. The attachments of both 3T3 and MDBK cells, especially to the fibronectin-immobilized swellable microbeads, were very high. Almost 96% of the 3T3 cells available in the cell culture medium did attach to these microbeads (2345 ± 98 cells per mg of polymer).