Hepatocyte immobilization on PHEMA microcarriers and its biologically modified forms.
Polyhydroxyethylmethacrylate (PHEMA) based microcarriers with different bulk structures were prepared by a phase inversion polymerization technique. PHEMA surfaces were further modified chemically by glow-discharge treatment, and biologically by covalent attachment of fibrinogen and collagen. Hepatocytes were isolated from young male Wistar rats using an in situ portal vein collagenase perfusion technique. Freshly isolated hepatocytes were seeded at 6 x 10(5) cells/mL and microcarrier concentration was 10 g/L. Stationary microcarrier cultures were carried out in standard (nontissue culture) polystyrene petri dishes in a humidified 5% CO2 incubator at 37 +/- 0.5 degrees C. Cell attachment was followed by light microscopy by taking samples from the culture medium every 30 min. Urea and protein syntheses by microcarrier-attached hepatocytes were determined by standard techniques. Nonswellable (highly cross-linked) hydrophilic PHEMA microcarriers did not support cell attachment and viability. However, swellable (low cross-linked) PHEMA microcarriers (pretreated in FBS) allowed high attachment and cell spreading. PHEMA microcarriers treated in dimethylaminoethylmethacrylate (DMAEMA) glow-discharge plasma also improved the cell attachment characteristics of the PHEMA microcarriers. The highest attachment efficiencies (immobilization yields) were observed with the biologically modified PHEMA microcarriers, especially modified with fibronectin. Metabolic activity, as estimated by urea and protein syntheses, was also higher in these microcarriers.
Liver Biosupport and Hepatitis Research Laboratory, UCLA School of Medicine 90024-7019.
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Dixit
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Piskin
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Arthur
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Denizli
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Tuncel
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Denkbas
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Gitnick
D000818:Animals; D001672:Biocompatible Materials; D002448:Cell Adhesion; D003094:Collagen; D046508:Culture Techniques / Q000379:methods; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D008855:Microscopy, Electron, Scanning; D010477:Perfusion; D011102:Polyhydroxyethyl Methacrylate; D011169:Portal Vein; D014176:Protein Biosynthesis; D051381:Rats; D017208:Rats, Wistar; D014508:Urea / Q000378:metabolism
D001672:Biocompatible Materials; D011102:Polyhydroxyethyl Methacrylate; D014508:Urea; D003094:Collagen