Rapidly Functional Immobilization of Immortalized Human Hepatocytes Using Cell Adhesive GRGDS Peptide-Carrying Cellulose Microspheres
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.
Document Type: Research Article
Affiliations: 1: *First Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan 2: †Second Department of Anatomy, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan 3: ‡Central Research Laboratory, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, Japan 4: §Department of Medical Products, Kuraray Co., Ltd., 1621 Sakazu, Kurashiki, Okayama, 710-8622 Japan
Publication date: April 1, 2001
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