title: Exogenous gene expression and growth regulation of hematopoietic cells via a novel human artificial chromosome.
abstract: A number of gene delivery systems are currently being developed for potential use in gene therapy. Here, we demonstrate the feasibility of 21deltaqHAC, a newly developed human artificial chromosome (HAC), as a gene delivery system. We first introduced a 21deltaqHAC carrying an EGFP reporter gene and a geneticin-resistant gene (EGFP-21deltaqHAC) into hematopoietic cells by microcell-mediated chromosome transfer. These HAC-containing hematopoietic cells showed resistance to geneticin, expressed EGFP and retained the ability to differentiate into various lineages, and the EGFP-21deltaqHAC was successfully transduced into primary hematopoietic cells. Hematopoietic cells harboring the EGFP-21deltaqHAC could still be detected at two weeks post-transplantation in immunodeficient mice. We also showed effective expansion of hematopoietic cells by introducing the 21deltaqHAC containing ScFvg, a gp130-based chimeric receptor that transmits growth signals in response to specific-antigen of this receptor. All of these results demonstrate the usefulness of HAC in gene therapy.
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan.|Department of Human Genome Science, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, 683-8503, Yonago, Tottori, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Yamada
authors.affiliation: Pharmaceutical Research Laboratories, Pharmaceutical Division, Kirin Brewery Co., Ltd, 3 Miyahara, Takasaki, Gunma 370-1295, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kunisato
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kawahara
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan.
authors.forename: Candice G T
authors.initials: CGT
authors.lastname: Tahimic
authors.affiliation: Department of Human Genome Science, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, 683-8503, Yonago, Tottori, Japan.
authors.forename: Xianying
authors.initials: X
authors.lastname: Ren
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ueda
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nagamune
authors.affiliation: Department of Human Genome Science, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, 683-8503, Yonago, Tottori, Japan.
authors.forename: Motonobu
authors.initials: M
authors.lastname: Katoh
authors.affiliation: Department of Human Genome Science, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, 683-8503, Yonago, Tottori, Japan.
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Inoue
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan.|Pharmaceutical Research Laboratories, Pharmaceutical Division, Kirin Brewery Co., Ltd, 3 Miyahara, Takasaki, Gunma 370-1295, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Nishikawa
authors.affiliation: Department of Biomedical Science, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, Yonago, Tottori 683-8503, Japan. oshimura@grape.med.tottori-u.ac.jp.|Department of Human Genome Science, Graduate School of Medical Science, Tottori University, 86 Nishi-cho, 683-8503, Yonago, Tottori, Japan. oshimura@grape.med.tottori-u.ac.jp.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Oshimura
mesh_terms: D000818:Animals; D022222:Chromosomes, Artificial, Human / Q000235:genetics*; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics*; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods*; D017931:DNA Primers; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D005839:Gentamicins; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D051379:Mice
chemical_list: D017931:DNA Primers; D005839:Gentamicins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C010680:antibiotic G 418