Rapidly functional immobilization of immortalized human hepatocytes using cell adhesive GRGDS peptide-carrying cellulose microspheres

N. Kobayashi, T. Taguchi, H. Noguchi, T. Okitsu, T. Totsugawa, T. Watanabe, T. Matsumura, T. Fujiwara, H. Urata, N. Kishimoto, N. Hayashi, S. Nakaji, T. Murakami, N. Tanaka

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)387-392
Number of pages6
JournalCell Transplantation
Volume10
Issue number4-5
DOIs
Publication statusPublished - 2001

Keywords

  • Artificial liver
  • Cellulose microspheres
  • Immortal hepatocytes

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

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