Coatings of low-density lipoprotein and synthetic glycoconjugates as substrata for hepatocytes.
Asialoglycoprotein (ASGP) receptors expressed on rat hepatocytes interact with glycoproteins containing galactose or N-acetylgalactosamine residues at the nonreducing termini of oligosaccharide chains to mediate endocytosis, and cholesterol transport protein with apolipoprotein B (LDL, low-density lipoprotein) in plasma interacts with LDL receptors and heparinoids in the extracellular matrix. We developed novel techniques to prepare galactose- and LDL-immobilized culture plates, using galactose-tagged polystyrene (galactose-carrying polystyrene [GalCPS]: N-p-vinylbenzyl-O-beta-D-galactopyranosyl-[1-->4]-D-gluconamide) and poly(2-acrylamide-2-methyl-1-propanesulfonate) (PAPS), respectively. Hepatocytes adhered well to plates coated with either GalCPS or LDL, and therefore the GalCPS- and LDL-coated plates were examined as specific substrata for culturing hepatocytes. These cultures promoted the formation of three-dimensional, multicellular aggregates with regulation of excess proliferation of non-parenchymal cells. Furthermore, the LDL coating resulted in higher albumin synthesis and an identical level of lactate dehydrogenase (LDH) compared with cells cultured on collagen- and GalCPS-coated plates. Thus, the two culture systems described here, and especially the LDL-coated plates, have potential for the development of a hybrid artificial liver.
Yura
Hirofumi
H
NeTech, Inc., Takatsu, Kawasaki, Kanagawa, Japan.
Ishihara
Masayuki
M
Nakamura
Shingo
S
Kishimoto
Satoko
S
Kanatani
Yasuhiro
Y
Horio
Takuya
T
Ishizuka
Takamitsu
T
Kawakami
Mitsuyuki
M
Matsui
Takemi
T
D000327:Adsorption; D000476:Alkanesulfonates / Q000737:chemistry; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D005690:Galactose / Q000737:chemistry / Q000494:pharmacology*; D006001:Glycoconjugates / Q000737:chemistry / Q000494:pharmacology*; D022781:Hepatocytes / Q000166:cytology* / Q000648:ultrastructure; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000494:pharmacology*; D019164:Liver, Artificial; D011137:Polystyrenes / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D000476:Alkanesulfonates; D006001:Glycoconjugates; D008077:Lipoproteins, LDL; D011137:Polystyrenes; D005690:Galactose