High density of immobilized galactose ligand enhances hepatocyte attachment and function.
Galactosylated surface is an attractive substrate for hepatocyte culture because of the specific interaction between the galactose ligand and the asialoglycoprotein receptor on hepatocytes. In this study, we described a scheme to achieve high density of immobilized galactose ligands on polyethylene terephthalate (PET) surface by first surface-grafting polyacrylic acid on plasma-pretreated PET film under UV irradiation, followed by conjugation of a galactose derivative (1-O-(6'-aminohexyl)-D-galactopyranoside) to the grafted polyacrylic acid chains. A high galactose density of 513 nmol/cm(2) on the PET surface was used in this study to investigate the behavior of cultured hepatocyte. This engineered substrate showed high affinity to fluorescein isothiocyanate-lectin binding. Primary rat hepatocytes, when seeded at a density of 2 x 10(5) cells/cm(2), attached to the galactosylated PET substrate at a similar efficiency compared with collagen-coated substrate. The hepatocytes spontaneously formed aggregates 1 day after cell seeding and showed better maintenance of albumin secretion and urea synthesis functions than those cultured on collagen-coated surface.
Yin
Chao
C
Tissue and Therapeutic Engineering Lab, Johns Hopkins Singapore, Level 5 Clinical Research Center, NUS, 10 Medical Drive, Singapore 117597.
Ying
Lei
L
Zhang
Peng-Chi
PC
Zhuo
Ren-Xi
RX
Kang
En-Tang
ET
Leong
Kam W
KW
Mao
Hai-Quan
HQ
D000180:Acrylic Resins / Q000737:chemistry; D000418:Albumins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D048429:Cell Size; D002478:Cells, Cultured; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D005690:Galactose / Q000737:chemistry* / Q000378:metabolism; D022781:Hepatocytes / Q000502:physiology* / Q000648:ultrastructure; D008024:Ligands; D015394:Molecular Structure; D037121:Plant Lectins / Q000378:metabolism; D011093:Polyethylene Terephthalates / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D013499:Surface Properties; D014508:Urea / Q000378:metabolism
D000180:Acrylic Resins; D000418:Albumins; D008024:Ligands; D037121:Plant Lectins; D011093:Polyethylene Terephthalates; C006903:carbopol 940; D014508:Urea; D016650:Fluorescein-5-isothiocyanate; D005690:Galactose