Digoxin transport by renal proximal tubule cells is enhanced by adhesive synthetic RGD peptide.
BACKGROUND
The dialyzer apparatus has been widely used as an artificial kidney in medical treatment. However, side effects such as amyloidosis have occurred during long-term treatment. Therefore, we focused on developing a hybrid artificial kidney with a filtration and reabsorption apparatus, but it was found that cells spread extensively and it is difficult to maintain a uniform monolayer with a regular cell shape on a collagen-coated substrate. The purpose of this study was to improve cell adhesion, uniform stable monolayer formation and active transport function by immobilization of arginine-glycine-aspartic acid (RGD) on the culture substratum.


METHODS
Polycarbonate semipermeable membranes were coated with collagen, fibronectin, laminin and synthetic polypeptide, including RGD (Pronectin F). Cell adhesion and digoxin transport were estimated using a renal proximal tubule cell line that overexpressed the P-glycoprotein gene.


CONCLUSIONS
Under initial and confluent conditions, immobilized cell density in Pronectin F-coated wells was higher than that under other conditions. Transepithelial electrical resistance and digoxin transport activity on Pronectin F-coated membranes were the highest of all conditions. This might have been caused by uniform cell morphology and high cell density.
Ijima
H
H
Department of Chemical Engineering, Faculty of Engineering, Graduate School, Kyushu University, Nishi-ku, Fukuoka, Japan. ijima@chem-eng.kyushu-u.ac.jp
Kuroda
S
S
Ono
T
T
Kawakami
K
K
D001693:Biological Transport, Active; D002448:Cell Adhesion / Q000187:drug effects*; D002452:Cell Count; D002460:Cell Line; D002478:Cells, Cultured; D024042:Collagen Type I / Q000494:pharmacology; D004058:Diffusion; D004077:Digoxin / Q000493:pharmacokinetics; D005353:Fibronectins / Q000494:pharmacology; D019450:Genes, MDR; D006801:Humans; D007444:Inulin / Q000493:pharmacokinetics; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology
D024042:Collagen Type I; D005353:Fibronectins; D009842:Oligopeptides; D011994:Recombinant Proteins; C070367:ProNectin F; D004077:Digoxin; C047981:arginyl-glycyl-aspartic acid; D007444:Inulin