RRC ID 12069
Author Kobayashi N, Taguchi T, Noguchi H, Okitsu T, Totsugawa T, Watanabe T, Matsumura T, Fujiwara T, Urata H, Kishimoto N, Hayashi N, Nakaji S, Murakami T, Tanaka N.
Title Rapidly functional immobilization of immortalized human hepatocytes using cell adhesive GRGDS peptide-carrying cellulose microspheres.
Journal Cell Transplant
Abstract With the development of biotechnology, hepatic support by a hybrid artificial liver (HAL) using hepatocytes has been given much attention. Because the availability of human livers is limited, we have established a tightly regulated immortal human hepatocyte cell line, NKNT-3, for developing HAL. Because high-density cell culture allows the compactness of the HAL device and its easy use under emergency circumstances, we have developed cell adhesive GRGDS peptide-containing cellulose microspheres (GRGDS/CMS). The GRGDS/CMS efficiently immobilized NKNT-3 cells within 24 h in a stirred suspension culture. Electron microscopic examinations demonstrated glycogen granules and well-developed endoplasmic reticulum and mitochondria in NKNT-3 cells attached to the GRGDS/CMS. The cells showed ammonia clearance activity, whereas HepG2-transformed human liver cells did not remove the loaded ammonia. An efficient adenoviral delivery of the lacZ reporter gene was performed in GRGDS/CMS-immobilized NKNT-3 cells. In this study we present rapid immobilization of NKNT-3 immortal human hepatocytes using cellulose microspheres carrying GRGDS peptides. These microspheres satisfied immediate preparation of NKNT-3 cells in sufficient quantity and of adequate quality.
Volume 10(4-5)
Pages 387-92
Published 2001-1-1
PMID 11549059
MeSH Ammonia / metabolism Cell Adhesion / physiology* Cell Culture Techniques / methods Cell Line Cell Size Cells, Immobilized* Cellulose / metabolism Gene Transfer Techniques Hepatocytes / physiology* Hepatocytes / ultrastructure Humans Liver, Artificial* Microspheres* Oligopeptides* / metabolism Time Factors
IF 3.341
Times Cited 11
DNA material AxCALacZ Adex1CAlacZ (RDB02726)