Updated on 2025/08/23

写真a

 
KUSAKABE Hirokazu
 
Organization
School of Medicine General Education Biology
External link

Degree

  • 博士(保健学) ( 1999.7   杏林大学 )

  • 修士(理学) ( 1992.3   弘前大学 )

Research Interests

  • spermatozoa

  • preservation

  • 配偶子

  • ゲノム

  • 精子

  • 保存

  • Gamete

  • Genome

Research Areas

  • Environmental Science/Agriculture Science / Environmental effects of radiation

  • Environmental Science/Agriculture Science / Biological resource conservation

  • Environmental Science/Agriculture Science / Environmental effects of chemicals

Education

  • 弘前大学大学院   理学研究科   生物学専攻

    1990.4 - 1992.3

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    Country: Japan

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  • Hirosaki University

    1986.4 - 1990.3

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    Country: Japan

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Research History

  • Asahikawa Medical College   Professor

    2024.4

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  • Asahikawa Medical College   School of Medicine   Associate Professor

    2012.4

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  • Asahikawa Medical College   School of Medicine   Lecturer

    2008.7 - 2012.3

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  • Asahikawa Medical College   School of Medicine   Assistant Professor

    2007.4 - 2008.6

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  • Asahikawa Medical College   Teaching Associate

    2002.4 - 2007.3

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  • Asahikawa Medical College   School of Medicine

    2002.4 - 2007.3

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  • Institute for Biogenesis Research   Asistant Reseracher

    1999.10 - 2001.10

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  • Assistant Researcher, Institute for Biogenesis

    1999 - 2001

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  • (財)食品薬品安全センター秦野研究所   研究技術員

    1992.4 - 1999.9

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  • Researcher, Food and Drug Safety Center/Hatano

    1992 - 1999

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Professional Memberships

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Studying abroad experiences

  • 1999.10 - 2001.9   Institute for Biogenesis Research, University of Hawaii at Manoa  

Papers

  • Chromosomal integrity of freeze-dried karyoplasts in mice. International journal

    Hirokazu Kusakabe

    Cryobiology   119   105247 - 105247   2025.6

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    Language:English   Publishing type:Research paper (scientific journal)  

    In mammalian species, there is currently no way to preserve mature oocytes at supra-zero temperatures without cryostorage. Metaphase II (MII) oocytes freeze-dried in any medium or solution cannot be revived after rehydration. Therefore, the injurious effects to chromosomes of freeze-drying MII oocytes have not been reported. The aim of this study was to examine the chromosomal integrity of freeze-dried MII oocytes in mice. Spindle apparatuses with small amounts of cytoplasm, "karyoplasts" so-called, were removed from MII mouse oocytes. Before freeze-drying (FD), the karyoplasts were incubated at 4 °C for up to 2 days (pre-FD incubation) in EGTA/Tris-HCl buffered solution supplemented with 20 μmol/l γ-tocotrienol. After freeze-drying, the freeze-dried karyoplasts were rehydrated and microinjected into enucleated MII oocytes. Parthenogenetic activation of the reconstructed oocytes was performed to analyze the chromosomes at the first cleavage metaphase. A portion of normally activated oocytes that had been injected with fresh karyoplasts (51 %) and karyoplasts freeze-dried after pre-FD incubation for 8 h to 2 d (27 %-29 %) exhibited normal chromosome constitution. Insufficient pre-FD incubation (0-4 h) caused severe chromosomal damage. By contrast, almost all parthenogenetic embryos (98 %) reconstructed via fusion of fresh karyoplasts and enucleated oocytes maintained normal chromosome constitution. Some MII oocytes reconstructed and activated parthenogenetically using freeze-dried karyoplasts could develop the first cleavage metaphase (67-80 %) while retaining chromosome stability (3-29 %). Further improvements in FD procedures should enhance the chromosomal integrity of freeze-dried karyoplasts.

    DOI: 10.1016/j.cryobiol.2025.105247

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  • A reliable technique for karyotyping mouse oocytes prepared by a gradual fixation/air-drying method followed by multicolour FISH. International journal

    Toshiaki Hino, Hirokazu Kusakabe

    Biology open   12 ( 12 )   2023.12

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    Language:English   Publishing type:Research paper (scientific journal)  

    Chromosome segregation errors during oocyte meiosis increase with age and lead to aneuploidy; hence, the mechanism has been studied extensively. The mouse is the most widely used experimental animal for this purpose. However, the lack of a reliable and efficient technique for karyotyping mouse oocytes has limited comprehensive studies of chromosome-specific segregation errors in this animal model. Here, we developed a novel karyotyping technique for mouse oocytes by applying multicolour fluorescence in situ hybridisation (FISH) to chromosome slides prepared by a gradual fixation/air-drying method, which is best suited to avoid rupture of oocyte membrane and artificial loss of chromosomes. The success rate of karyotyping meiosis I and II oocytes was about 30%, which improved to over 90% when the oocytes were 'flattened' during fixation and the chromosome specimens were denatured at 4°C. When this technique was applied to the karyotyping of meiosis II oocytes from aged female mice and from young female mice injected with colchicine, more than 80% of the oocytes were successfully karyotyped and the number of chromosomes was identified on all aberrant chromosomes. In conclusion, our technique allows for the efficient and reliable karyotyping of mouse oocytes.

    DOI: 10.1242/bio.060188

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  • Sperm acrosome status before and during fertilization in the Chinese hamster ( Cricetulus griseus ), and observation of oviductal vesicles and globules Reviewed

    Hiroyuki Tateno, Miwa Tamura‐Nakano, Hirokazu Kusakabe, Noritaka Hirohashi, Natsuko Kawano, Ryuzo Yanagimachi

    Molecular Reproduction and Development   88 ( 12 )   793 - 804   2021.12

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    DOI: 10.1002/mrd.23547

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1002/mrd.23547

  • Production of mouse fetuses using spermatozoa exposed temporarily to high temperature or continuously to room temperature after freeze-drying in Na+-free/K+-rich EGTA buffer Reviewed

    Hirokazu Kusakabe

    Cryobiology   87   105 - 109   2019.4

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  • Prevention of high-temperature-induced chromosome damage in mouse spermatozoa freeze-dried using Ca2+ chelator-containing buffer alkalinized with NaOH or KOH Reviewed

    Hirokazu Kusakabe, Hiroyuki Tateno

    CRYOBIOLOGY   79   71 - 77   2017.12

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/j.cryobiol.2017.08.007

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  • マウス第二極体ゲノムに由来する混倍数性受精卵のゲノム動態とその発生能

    日野 敏昭, 日下部 博一, 立野 裕幸

    Journal of Mammalian Ova Research   31 ( 2 )   S59 - S59   2014.4

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    Language:Japanese   Publisher:(一社)日本卵子学会  

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  • マウス第二極体に由来する混倍数性受精卵のゲノム動態

    日野 敏昭, 日下部 博一, 立野 裕幸

    日本生殖医学会雑誌   58 ( 4 )   378 - 378   2013.10

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    Language:Japanese   Publisher:(一社)日本生殖医学会  

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  • Production of offspring after sperm chromosome screening: an experiment using the mouse model Reviewed

    H. Watanabe, H. Kusakabe, H. Mori, R. Yanagimachi, H. Tateno

    HUMAN REPRODUCTION   28 ( 2 )   531 - 537   2013.2

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    DOI: 10.1093/humrep/des388

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  • Chromosomal stability of second polar bodies in mouse embryos Reviewed

    Toshiaki Hino, Hirokazu Kusakabe, Hiroyuki Tateno

    JOURNAL OF ASSISTED REPRODUCTION AND GENETICS   30 ( 1 )   91 - 98   2013.1

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER/PLENUM PUBLISHERS  

    Incorporation of a second polar body (PB2) into one of the blastomeres has been considered as a causal mechanism underlying diploid/triploid mixoploidy in humans. Using a mouse model, we examined whether PB2s can participate in the formation of mixoploidy.
    Uptake of BrdU was examined to determine DNA synthesis in PB2s up to 28 h after fertilization. PB2s from embryos at 4-6 (1-cell), 24 (2-cell), 48 (4-cell), and 72 h (morula) were fused with MII oocytes to induce premature chromosome condensation. Caspase and TUNEL assays were used to detect apoptotic PB2s at 24, 48, and 72 h. PB2s were fused with one of the blastomeres of the 2-cell embryos to produce mixoploid embryos.
    DNA synthesis in the PB2s continued until 22 h after fertilization. At 4-6 h, nearly all of the PB2s showed G1-type chromosomes and there was no significant increase in chromosome damage. At 24, 48, and 72 h, S-type chromatin predominated. Few PB2s showed apoptotic response until 72 h. Regardless of the fusion with the PB2, more than 90 % of the embryos developed to 4-cell stage, and over 80 % of the resultant 4-cell embryos had daughter blastomeres with a morphologically normal nucleus. Some of the daughter blastomeres displayed triploidy.
    The PB2 is viable for at least 72 h after fertilization, with slow progression through the cell cycle. Once the PB2 has been incorporated into a blastomere, the cell cycle of the PB2 might be synchronized with that of the host resulting in diploid/triploid mixoploidy.

    DOI: 10.1007/s10815-012-9899-3

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  • マウスおよびヒト凍結乾燥精子ゲノムの高温耐性獲得に関する研究

    日下部 博一

    旭川医科大学研究フォーラム   13 ( 1 )   86 - 88   2013

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    Authorship:Lead author   Language:Japanese   Publishing type:Research paper (bulletin of university, research institution)  

    雑誌掲載版

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    Other Link: http://search.jamas.or.jp/link/ui/2013338948

  • Chromosomal stability of second polar bodies Reviewed

    Hino T, Kusakabe H, Tateno H

    J Assisted Reprod Genet   30   91 - 98   2013

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    DOI: 10.1007/s10815-012-9899-3

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  • Characterization of chromosomal damage accumulated in freeze-dried mouse spermatozoa preserved under ambient and heat stress conditions Reviewed

    Hirokazu Kusakabe, Hiroyuki Tateno

    MUTAGENESIS   26 ( 3 )   447 - 453   2011.5

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/mutage/ger003

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  • Structural chromosomal aberrations, aneuploidy, and mosaicism in early cleavage mouse embryos derived from spermatozoa exposed to gamma-rays Reviewed

    Hiroyuki Tateno, Hirokazu Kusakabe, Yujiroh Kamiguchi

    INTERNATIONAL JOURNAL OF RADIATION BIOLOGY   87 ( 3 )   320 - 329   2011.3

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.3109/09553002.2011.530334

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  • Shortening of alkaline DNA unwinding time does not interfere with detecting DNA damage to mouse and human spermatozoa in the comet assay Reviewed

    Hirokazu Kusakabe, Hiroyuki Tateno

    ASIAN JOURNAL OF ANDROLOGY   13 ( 1 )   172 - 174   2011.1

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1038/aja.2010.105

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  • Chromosomal integrity and DNA damage in freeze-dried spermatozoa Invited Reviewed

    Hirokazu Kusakabe

    Reproductive Medicine and Biology   10 ( 4 )   199 - 210   2011

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (conference, symposium, etc.)   Publisher:John Wiley and Sons Ltd  

    DOI: 10.1007/s12522-011-0092-7

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  • Varidity of a simple vitrification technique for chromosome study of mouse one-cell embryos. Reviewed

    Hino T, Kusakabe H, Tateno H

    Chrom Sci   13 ( 3 )   45 - 48   2010

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:THE SOCIETY OF CHROMOSOME RESEARCH  

    To evaluate cytogenetic validity of a simple vitrification technique for embryo cryopreservation, mouse 1-cell embryos were vitrified 4, 6, and 8 h after <i>in vitro</i> fertilization (IVF). In addition, chromosomal damage of spermatozoa treated with methyl methanesulfonate (MMS) was estimated using vitrified 1-cell embryos. More than 90% of embryos survived vitrification regardless of the time after IVF. In the 4-h and 6-h groups, some of the surviving embryos swelled after recovery. The incidence of structural chromosome aberrations and aneuploidy in embryos with morphologically normal features did not significantly increase in any group. The vitrification technique preserved 1-cell embryos derived from MMS-treated spermatozoa without alteration of chromosome damage. This technique will enable us to manage the optimal time for chromosome preparation of mouse 1-cell embryos.

    DOI: 10.11352/scr.13.45

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    Other Link: http://amcor.asahikawa-med.ac.jp/modules/xoonips/detail.php?id=CS13-3-45

  • マウス精子を用いる単一細胞ゲル電気泳動法(コメットアッセイ)および染色体分析法による遺伝的傷害の検出感度比較.

    日下部 博一

    旭川医科大学研究フォーラム   9 ( 1 )   77 - 78   2009

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    Authorship:Lead author   Language:Japanese   Publishing type:Research paper (bulletin of university, research institution)  

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    Other Link: http://search.jamas.or.jp/link/ui/2009233001

  • Mouse and human spermatozoa can be freeze-dried without damaging their chromosomes (in Russian)

    Kusakabe H., Yanagimachi R., Kamiguchi Y.

    Hum Reprod (Specialized Edition, in Russian)   5   82 - 88   2008.4

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  • Mouse and human spermatozoa can be freeze-dried without damaging their chromosomes Reviewed

    H. Kusakabe, R. Yanagimachi, Y. Kamiguchi

    HUMAN REPRODUCTION   23 ( 2 )   233 - 239   2008.2

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1093/humrep/dem252

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  • Fertilizability and chromosomal integrity of frozen-thawed Bryde&apos;s whale (Balaenoptera edeni) spermatozoa intracytoplasmically injected into mouse oocytes Reviewed

    H. Watanabe, H. Tateno, H. Kusakabe, T. Matsuoka, Y. Kamiguchi, Y. Fujise, H. Ishikawa, S. Ohsumi, Y. Fukui

    ZYGOTE   15 ( 1 )   9 - 14   2007.2

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1017/S0967199406003923

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  • Chromosome analysis of mouse zygotes after injecting oocytes with spermatozoa treated in vitro with green tea catechin, (-)-epigallocatechin gallate (EGCG). Reviewed

    Kusakabe H, Kamiguchi Y

    Mutation research   564 ( 2 )   195 - 200   2004.12

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    DOI: 10.1016/j.mrgentox.2004.08.005

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  • Chromosomal integrity of freeze-dried mouse spermatozoa after Cs-137 gamma-ray irradiation Reviewed

    Kusakabe H, Kamiguchi Y

    MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS   556 ( 1-2 )   163 - 168   2004.11

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    DOI: 10.1016/j.mrfmmm.2004.08.001

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  • Ability to activate oocytes and chromosome integrity of mouse spermatozoa preserved in EGTA Tris-HCl buffered solution supplemented with antioxidants Reviewed

    Kusakabe H, Kamiguchi Y

    THERIOGENOLOGY   62 ( 5 )   897 - 905   2004.9

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    DOI: 10.1016/j.theriogenology.2003.12.008

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  • Freeze-dried sperm fertilization leads to full-term development in rabbits Reviewed

    JL Liu, H Kusakabe, CC Chang, H Suzuki, DW Schmidt, M Julian, R Pfeffer, CL Bormann, XC Tian, R Yanagimachi, XZ Yang

    BIOLOGY OF REPRODUCTION   70 ( 6 )   1776 - 1781   2004.6

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    DOI: 10.1095/biolreprod.103.025957

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  • Long-term preservation of mouse spermatozoa after freeze-drying and freezing without cryoprotection Reviewed

    MA Ward, T Kaneko, H Kusakabe, JD Biggers, DG Whittingham, R Yanagimachi

    BIOLOGY OF REPRODUCTION   69 ( 6 )   2100 - 2108   2003.12

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    DOI: 10.1095/biolreprod.103.020529

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  • Analysis of sperm chromosomes in infertile males with teratozoospermia. (Polish) Reviewed

    Szczygiel MA, Kusakabe H, Wiland E, Kurpisz M

    Ginekol Pol   74   108 - 115   2003

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  • Intracytoplasmic sperm injection is more efficient than in vitro fertilization for generating mouse embryos from cryopreserved spermatozoa Reviewed

    MA Szczygiel, H Kusakabe, R Yanagimachi, DG Whittingham

    BIOLOGY OF REPRODUCTION   67 ( 4 )   1278 - 1284   2002.10

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    DOI: 10.1095/biolreprod.102.006106

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  • Separation of motile populations of spermatozoa prior to freezing is beneficial for subsequent fertilization in vitro: A study with various mouse strains Reviewed

    MA Szczygiel, H Kusakabe, R Yanagimachi, DG Whittingham

    BIOLOGY OF REPRODUCTION   67 ( 1 )   287 - 292   2002.7

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  • Relevance of chemical structure and cytotoxicity to the induction of chromosome aberrations based on the testing results of 98 high production volume industrial chemicals Reviewed

    H Kusakabe, K Yamakage, S Wakuri, K Sasaki, Y Nakagawa, M Watanabe, M Hayashi, T Sofuni, H Ono, N Tanaka

    MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS   517 ( 1-2 )   187 - 198   2002.5

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1016/S1383-5718(02)00062-1

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  • マウス凍結乾燥精子の卵細胞質内精子注入法(ICSI法)

    日下部 博一

    秦野研究所年報   25   102 - 108   2002

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  • Adult murine neurons: their chromatin and chromosome changes and failure to support embryonic development as revealed by nuclear transfer Reviewed

    T Osada, H Kusakabe, H Akutsu, T Yagi, R Yanagimachi

    CYTOGENETIC AND GENOME RESEARCH   97 ( 1-2 )   7 - 12   2002

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    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1159/000064037

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  • Maintenance of genetic integrity in frozen and freeze-dried mouse spermatozoa Reviewed

    H Kusakabe, MA Szczygiel, DG Whittingham, R Yanagimachi

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   98 ( 24 )   13501 - 13506   2001.11

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1073/pnas.241517598

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  • ヒト体細胞および精子における変異原物質誘発染色体転座に関する研究 Reviewed

    日下部博一

    杏林医学会雑誌   30 ( 4 )   573 - 574   1999.9

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    Authorship:Lead author   Language:Japanese   Publishing type:Doctoral thesis   Publisher:杏林医学会  

    DOI: 10.11434/kyorinmed.30.573

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  • Chromosome-type aberrations induced in chromosome 9 after treatment of human peripheral blood lymphocytes with mitomycin C at the G(0) phase Reviewed

    H Kusakabe, T Takahashi, N Tanaka

    CYTOGENETICS AND CELL GENETICS   85 ( 3-4 )   212 - 216   1999

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  • Comparative studies of MCL-5 cells and human lymphocytes for detecting indirect-acting clastogens Reviewed

    K Yamakage, H Kusakabe, N Tanaka

    MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS   412 ( 1 )   55 - 61   1998.1

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    DOI: 10.1016/S1383-5718(97)00170-8

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  • Re-evaluation of chromosomal aberration induction on nine mouse lymphoma assay 'unique positive' NTP carcinogens Reviewed

    A Matsuoka, K Yamakage, H Kusakabe, S Wakuri, M Asakura, T Noguchi, T Sugiyama, H Shimada, S Nakayama, Y Kasahara, Y Takahashi, KF Miura, M Hatanaka, M Ishidate, T Morita, K Watanabe, M Hara, K Odawara, N Tanaka, M Hayashi, T Sofuni

    MUTATION RESEARCH-GENETIC TOXICOLOGY   369 ( 3-4 )   243 - 252   1996.8

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  • Detection of neocarzinostatin-induced translocations in human sperm chromosomes using fluorescence in situ hybridization of chromosome 2 Reviewed

    H Kusakabe, K Yamakage, N Tanaka

    MUTATION RESEARCH-GENETIC TOXICOLOGY   369 ( 1-2 )   51 - 58   1996.7

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    DOI: 10.1016/S0165-1218(96)90047-6

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  • Revised karyotypes of the Japanese Northern Red-backed Vole, Clethrionomys rutilus mikado. Reviewed

    Obara Y, Kusakabe H, Miyakoshi K, Kawada S

    J Mamm Soc Japan   20   125 - 133   1995

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    DOI: 10.11238/jmammsocjapan.20.125

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  • Cytogenetic study on the adriamycin-induced Robertsonian fusion in three species of Clethrionomys Reviewed

    Hirokazu Kusakabe

    1992.3

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MISC

  • FOR HOW LONG DOES THE SECOND POLAR BODY OF THE MOUSE-FERTILIZED EGG RETAIN GENETIC STABILITY?

    T. Hino, H. Kusakabe, H. Tateno

    FERTILITY AND STERILITY   98 ( 3 )   S163 - S163   2012.9

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  • Occurrence of structural chromosome aberrations, aneuploidy and mosaicism in mouse early cleavage embryos derived from spermatozoa with γ-ray-induced DNA damage

    TATENO Hiroyuki, KUSAKABE Hirokazu, KAMIGUCHI Yujiroh

    28 ( 2 )   S112   2011.4

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    Language:Japanese  

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  • Fertilizability and chromosomal integrity of frozen-thawed bryde's whale (Balaenoptera edeni) spermatozoa intracytoplasmically injected into mouse oocytes

    H. Watanabe, H. Tateno, H. Kusakabe, Y. Kamiguchi, Y. Fujise, H. Ishikawa, S. Ohsumi, Y. Fukui

    REPRODUCTION FERTILITY AND DEVELOPMENT   19 ( 1 )   306 - 306   2007

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  • O-16. Novel method to improve chromosomal integrity of freeze-dried mouse spermatozoa(Abstracts of the oral and poster presentations) :

    KUSAKABE Hirokazu, KAMIGUCHI Yujiroh

    Chromosome science   9 ( 4 )   119 - 119   2006

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    Language:English   Publisher:Society of Chromosome Research  

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    Other Link: http://id.nii.ac.jp/1141/00107646/

  • Freeze-dried sperm fertilization leads to full-term development in rabbits

    JL Liu, H Kusakabe, CC Chang, H Suzuki, DW Schmidt, M Julian, R Pfeffer, CL Bormann, XC Tian, R Yanagimachi, XZ Yang

    BIOLOGY OF REPRODUCTION   70 ( 6 )   1776 - 1781   2004.6

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  • Long-term preservation of mouse spermatozoa after freeze-drying and freezing without cryoprotection

    MA Ward, T Kaneko, H Kusakabe, JD Biggers, DG Whittingham, R Yanagimachi

    BIOLOGY OF REPRODUCTION   69 ( 6 )   2100 - 2108   2003.12

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  • Long-term preservation of freeze-dried and frozen without cryoprotection mouse spermatozoa.

    MA Szczygiel, T Kaneko, H Kusakabe, DG Whittingham, R Yanagimachi

    BIOLOGY OF REPRODUCTION   68   209 - 209   2003

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  • Analysis of sperm chromosomes in inferile males with teratozoospermia"jointly worked"

    Gynekol Pol.   74, 108-115   2003

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  • Intracytoplasmic sperm injection is more efficient than in vitro fertilization for cryopreserved spermatozoa.

    MA Szczygiel, H Kusakabe, DG Whittingham, R Yanagimachi

    BIOLOGY OF REPRODUCTION   66   196 - 196   2002

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  • Adult murine neurons: their chromatin and chromosome changes and failure to support embryonic development as revealed by nuclear transfer

    T Osada, H Kusakabe, H Akutsu, T Yagi, R Yanagimachi

    CYTOGENETIC AND GENOME RESEARCH   97 ( 1-2 )   7 - 12   2002

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  • Comparison of the development of andrognetic and parthenogenetic mouse embryos.

    H Akutsu, H Tateno, H Kusakabe, R Yanagimachi

    BIOLOGY OF REPRODUCTION   62   253 - 254   2000

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    Web of Science

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Presentations

  • マウス凍結乾燥ミニ卵の作製と染色体ダメージについて

    日下部博一

    第69回日本生殖医学会学術講演会・総会  2024.11 

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    Event date: 2024.11

    Language:Japanese   Presentation type:Poster presentation  

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  • マウス凍結乾燥ミニ卵の γ - トコトリエノール処理による染色体ダメージの軽減

    日下部博一

    日本環境変異原ゲノム学会 

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    Event date: 2023.11

    Language:Japanese   Presentation type:Poster presentation  

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  • フリーズドライ精子作製用の精子処理液のpHと精子染色体異常の誘発について

    日下部博一, 立野裕幸

    財団法人染色体学会第65回(2014年度)年会  財団法人染色体学会

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    Event date: 2014.10

    Language:Japanese   Presentation type:Poster presentation  

    Venue:倉敷市  

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  • 室温保存したマウス凍結乾燥精子の染色体正常性

    日下部 博一, 上口勇次郎

    第51回日本不妊学会北海道地方部会・学術講演会 

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    Event date: 2005.2

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:札幌  

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  • 「生殖医療および遺伝子資源保存のための基礎研究」

    日下部 博一

    検査技術科学特別講演 

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    Event date: 2004.4

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:弘前  

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  • Maintenance of genetic integrity in frozen and freeze-dried mouse spermatozoa. Invited International conference

    Kusakabe H, Szczygiel MA, Whittingham DG, Yanagimachi R

    Sperm Club Meeting (National Cooperative Program on Mouse Sperm Cryopreservation)  2001.7 

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  • 室温・高温保存したフリーズドライ精子における染色体異常誘発の抑制 Invited

    日下部 博一

    日本卵子学会  2016.5 

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    Presentation type:Symposium, workshop panel (nominated)  

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  • キレート剤を使用するマウス精子の凍結乾燥保存法とゲノム正常性の維持

    日下部博一, Szczygiel MA, Whittingham DG, 柳町隆造

    日本不妊学会北海道地方部会総会・学術講演会  2003.3 

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  • 凍結乾燥処理後のマウス精子における染色体正常性の維持について

    日下部博一, Szczygiel MA, Whittingham DG, 柳町隆造

    染色体学会第53回大会  2002.10 

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  • 抗酸化物質を含む凍結乾燥用溶液中で室温保存したマウス精子の正常性

    日下部博一, 上口勇次郎

    日本不妊学会北海道地方部会総会・学術講演会  2004.3 

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  • ビタミンEで処理した非凍結保存精子の卵活性化能および染色体正常性について

    日下部博一, 上口勇次郎

    日本環境変異原学会第32回大会  2003.11 

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  • 生殖医療および遺伝子資源保存のための基礎研究 Invited

    日下部博一

    検査技術科学特別講演  2004.6 

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  • ガンマ線照射された凍結乾燥精子の染色体正常性

    日下部博一, 上口勇次郎

    日本環境変異原学会第33回大会  2004.11 

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  • 緑茶カテキンで処理されたマウス精子の染色体分析

    日下部博一, 上口勇次郎

    染色体学会第55回大会  2004.11 

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  • Separation of motile populations of mouse spermatozoa from various inbred and hybrid mice. International conference

    Szczygiel MA, Kusakabe H, Whittingham DG, Yanagimachi R

    Annual Meeting of the Society for Reproduction and Fertility  2001.11 

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  • A novel medium for the maintenance of chromosome integrity in freeze-dried mousespermatozoa. International conference

    Kusakabe H, Szczygiel MA, Whittingham DG, Yanagimachi R

    Annual Meeting of the Society for Reproduction and Fertility  2001.11 

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  • マウスにおける精子凍結乾燥法の改良と受精卵の染色体分析結果が示す同法の特徴

    日下部博一, 上口勇次郎

    染色体学会第57回大会  2006.11 

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  • 室温保存したマウス凍結乾燥精子の染色体正常性

    日下部博一, 上口勇次郎

    日本不妊学会北海道地方部会総会・学術講演会  2005.11 

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  • 高温保存した凍結乾燥精子の単一細胞ゲル電気泳動法(コメットアッセイ)によるDNA損傷の検出

    日下部博一

    第51回日本不妊学会北海道地方部会・学術講演会  2009.2 

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  • マウスおよびヒト精子または凍結乾燥精子を用いるコメットアッセイのDNA傷害検出能について

    日下部博一

    日本環境変異原学会第回37回大会  2008.12 

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  • 凍結乾燥精子の高温耐性獲得と染色体異常の誘発抑制

    日下部博一, 立野裕幸

    染色体学会第62回大会  2011.11 

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    Language:Chinese   Presentation type:Poster presentation  

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  • 高温(50℃)で保存した凍結乾燥精子におけるDNA損傷と染色分体交換の高頻度誘発

    日下部博一

    日本環境変異原学会第回37回大会  2009.11 

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  • フリーズドライ精子における染色体異常生成とその抑制 Invited

    日下部博一

    第58回日本生殖医学会  2013.11 

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    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

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  • 冷蔵および室温保存したマウス凍結乾燥精子における染色体分析

    日下部博一, 上口勇次郎

    染色体学会第56回大会  2005.10 

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  • Chromosomal damage in motile and immotile mouse spermatozoa treated in vitro with green tea catechin, (-)-epigallocatechin gallate (EGCG) International conference

    Kusakabe H, Kamiguchi Y

    9th International Conference on Environmental Mutagens  2005.9 

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  • 照射コムギ粉飼料給餌によるラット骨髄細胞と末梢血中における数的異常誘発

    田中憲穂, 山影康次, 若栗忍, 日下部博一

    日本環境変異原学会第21回大会  1992.11 

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  • ヤチネズミ属3 種のアドリアマイシン誘導ロバートソン型動原体融合について

    日下部博一, 小原良孝

    日本動物学会東北支部大会  1992.7 

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  • 染色体バンディングおよび FISH 法による染色体異常の継代変化の推移

    日下部博一, 田中憲穂

    日本環境変異原学会第22回大会  1993.11 

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  • マウス末梢血を用いた小核試験による染色体の数的異常誘発物質の検出

    山影康次, 若栗忍, 日下部博一, 田中憲穂

    日本環境変異原学会第21回大会  1992.11 

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  • マウス末梢血を用いた小核試験による染色体の数的異常誘発物質の検出

    山影康次, 中川ゆづき, 若栗忍, 日下部博一, 坂本京子, 澁谷徹, 西村茂, 奥苑正光, 田中憲穂

    日本環境変異原学会第24回大会  1995.11 

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  • 第2 染色体ペインティングによる化学物質誘発ヒト精子染色体転座の検出

    日下部博一, 立野裕幸, 上口勇次郎, 田中憲穂

    日本環境変異原学会第23回大会  1994.11 

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  • 抗癌剤ネオカルチノスタチン(NCS)の転座誘発

    日下部博一, 宮下めぐみ, 山影康次, 田中憲穂

    染色体学会第48回大会  1997.9 

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  • FISHを用いたヒトリンパ球異数性細胞の検出

    日下部博一, 山影康次, 若栗忍, 中川ゆづき, 田中憲穂

    日本環境変異原学会第24回大会  1995.11 

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  • マイトマイシン誘発ヒトリンパ球染色体転座について

    日下部博一, 宮下めぐみ, 井上みゆき, 高橋俊孝, 田中憲穂

    日本環境変異原学会第26回大会  1997.12 

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  • Neocarzinostatin-induced chromosome translocations in human sperm and peripheral blood lymphocytes International conference

    Kusakabe H, Yamakage K, Tanaka N

    7th International Conference on Environmental Mutagens  1997.9 

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  • FISH によるマイトマイシンC誘発ヒト9番染色体異常の分析

    日下部博一, 井上みゆき, 高橋俊孝, 田中憲穂

    染色体学会第49回大会  1998.11 

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  • 110 種の既存化学物質における染色体異常試験結果の総括および問題点

    日下部博一, 山影康次, 若栗 忍, 高橋俊孝, 佐々木澄志, 中川ゆづき, 井上みゆき, 渡辺美香, 橋本恵子, 田中憲穂

    日本環境変異原学会第27回大会  1998.11 

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  • L5178Y tk+/- 細胞を用いた突然変異と染色体異常誘発の予測

    若栗忍, 山影康次, 日下部博一, 吉永紀久子, 田中憲穂

    日本環境変異原学会第23回大会  1994 

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Industrial property rights

  • 哺乳類由来の細胞又は精子の凍結乾燥用溶液

    日下部博一

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    Application no:特願2006-080682  Date applied:2006.3

    Patent/Registration no:特許4967119号  Date registered:2012.4 

    Country of applicant:Domestic  

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  • 哺乳類由来の細胞又は精子の凍結乾燥用溶液

    日下部 博一

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    Application no:特願2006-080682  Date applied:2006.3

    Patent/Registration no:特許4967119  Date issued:2012.4

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Awards

  • 最優秀演題賞

    2009.2   第51回日本不妊学会北海道地方部会   高温保存した凍結乾燥精子の単一細胞ゲル電気泳動法(コメットアッセイ)によるDNA損傷の検出

    日下部 博一

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  • 第51回日本生殖医学会北海道地方部会 優秀演題賞

    2009.2   日本生殖医学会  

    日下部博一

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    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

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Research Projects

  • 凍結乾燥ミニ卵を用いるミトコンドリア置換法の安全性向上に関する研究

    2024.4 - 2027.3

    日本学術振興会  科学研究費助成事業 基盤研究(C) 

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    ミトコンドリア病をもたらす変異遺伝子を次世代に持ち込ませないための生殖医療技術「ミトコンドリア置換法」に、凍結乾燥卵(凍結乾燥ミニ卵)を使用することが可能かどうかを検討する。

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  • Freeze-dying of mammalian oocytes

    Grant number:20K06457  2020.4 - 2023.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

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    Grant amount:\3,770,000 ( Direct Cost: \2,900,000 、 Indirect Cost:\870,000 )

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  • 哺乳類のフリーズドライ精子における高温耐性獲得に関する研究

    2013.4 - 2016.3

    日本学術振興会  科学研究費補助金(基盤研究C) 

    日下部 博一

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    Authorship:Principal investigator  Grant type:Competitive

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  • マウスおよびヒト凍結乾燥精子ゲノムの高温耐性獲得に関する研究

    2011.10 - 2012.10

    旭川医科大学 

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  • マウス精子を用いる単一細胞ゲル電気泳動法(コメットアッセイ)および染色体分析法による遺伝的傷害の検出感度比較

    2007.10 - 2008.10

    旭川医科大学 

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  • マウス精子および体細胞の室温保存法に関する基礎的な研究

    Grant number:16681020  2004.4 - 2007.3

    日本学術振興会  科学研究費助成事業  若手研究(A)

    日下部 博一

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    Grant amount:\7,540,000 ( Direct Cost: \5,800,000 、 Indirect Cost:\1,740,000 )

    昨年までの研究により、新しいマウス精子凍結乾燥用溶液として考案した修正型ETBS(50 mMEGTA、100mM Tris-HCl(pH 7.4))に精子を前処理(プレインキュベーション、4℃または25℃で4〜7日間)することにより、凍結乾燥精子の染色体正常率が70-90%と高くなり、その受精卵の発生能は1年間冷蔵しても変わらず、また1年間室温保存した精子からも胎仔が得られることもわかっている。研究の最終年度である本年度は、この修正型凍結乾燥溶液の適用範囲を調べるため、健常なヒトの精子を予め37℃で保温した修正型ETBSに加え、スイムアップ(10分間)後に回収し凍結乾燥した。1ヶ月以内で冷蔵した凍結乾燥精子をマウスの除核卵に注入し、精子染色体を形成させてその染色体異常の頻度を調べた。その結果、マウスの場合とは異なり、ヒト精子を凍結乾燥する場合はプレインキュベーション時間をとらなくても染色体正常率が86-95%と十分に高いことから、凍結乾燥後の精子染色体の正常性を維持するためのプレインキュベーション時間の長さは種によって異なることが示唆された。加えて通常の染色レベルで正常と判断された20個の精子染色体像をランダムに選び、キナクリンマスタード染色によるQバンド分析も行ったところ、全ての凍結乾燥精子の染色体において転座、欠失などの異常は認められなかった。更に、精子が除核卵の中で中期染色体を形成できる頻度は新鮮精子(非凍結)よりも凍結乾燥精子の方が圧倒的に高く、新鮮精子が5-64%であったのに対し、凍結乾燥精子では86-92%(プレインキュベーション時間:4時間以内)となった。ヒト凍結乾燥精子の染色体正常性の維持とマウス凍結乾燥精子の発生能の高さは、本法が大型哺乳類の精液サンプルに対しても安全かつ対応可能であることを示唆しており、本結果ついては現在論文投稿中である。

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  • Study on reproductive technology to maintain the genetic integrity of freeze-dried spermatozoa and somatic cells in mammalian species

    2004

    Hirokazu Kusakabe

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  • Genotoxicity test of food additives

    2002.4 - 2003.3

    Food and Drug Safety Center 

    Hirokazu Kusakabe

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  • Changeable aspects of radiation-induced chromosome aberrations during early development of the zygote

    Grant number:13680615  2001 - 2004

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)

    KAMIGUCHI Yujiroh, TATENO Hiroyuki, KUSAKABE Hirokazu

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    Grant amount:\3,700,000 ( Direct Cost: \3,700,000 )

    To understand changeable aspects of radiation-induced chromosome aberrations (CAs) during the embryonic development, mouse pie-implantation and post-implantation embryos derived from gamma-irradiated (2,4 Gy) spermatozoa were studied cytogenetically and embryologically.
    1.Ahighly successful method for chmmosome preparation of mouse 1-cell to 4-cell embryos was established.
    2.A total number of 1,170 zygotes were karyotyped in irradiated and control groups. The zygotes with structural chromosome aberrations (SCAs) showed a clear dose-dependent increase, reaching 36% in the 4 Gy group. No dose-dependent increase of aneuploidy was found. The most predominant type of CA was chromosome breaks/fragments (60%) which was followed by dicentric chromosomes (25%).
    3.A total number of 396 2-cell embryos were karyotyped. The incidence of embryos with SCAs was nearly the same as that in 1-cell embryos, but about 90% of them converted into mosaic-type SCA. The incidence of SCA-derived aneuploidy was also significantly high in the irradiated groups.
    4.A total number of 537 4-cell embryos were karyotyped. A great majority of the embryos with SCA were of mosaic-type. The incidence of aneuploidy became decreased, suggesting that they converted again into mosaic type.
    5.About 50 to 60% of embryos with SCA reached to blastocyst stage, showing no developmental arrest or delay.
    6.Chromosome analysis was carried out with 2,240 somatic cells obtained from 85 live embryos (10〜11-day old). None of embryos was found to be chromosomally abnormal. The incidence of post-implantation death was significantly high in the irradiated groups, almost all the dead embryos were being in the state of degenerated conceptus. These suggest that selection of the embryos with CA mainly takes place immediately after implantation.
    7.No increase of embryonic death or developmental anomalies was observed in 16〜17-day-old fetuses (survey of 52 pregnant females), supporting the above-mentioned suggestion.

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Other

  • 【メディア報道】マウスの凍結乾燥精子の染色体正常率を改善し、世界で初めてヒトの凍結乾燥精子の染色体分析を行った研究内容が、2007年12月12日の読売新聞(夕刊)と翌日12月13日の北海道新聞(朝刊)の記事として紹介された。

    2007.12

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Social Activities

  • 旭川ウェルビーイング・コンソーシアム連携公開講座等事業部会委員

    2023.4

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    「あさひかわオープンカレッジ」における講演者の選出と依頼

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  • 令和2年度HOKKAIDOサイエンスフェスティバル

    2021.1 - 2021.2

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    北海道地区のSSH指定校の生徒が,各校における活動状況や研究成果の発表(ypou tube限定公開)を行い議論することで,相互に刺激し合い,研究内容の深化や研究活動の活性化を図るものである。

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  • 平成30年度北海道旭川西高等学校「SSH課題研究中間報告会」および「SSH課題研究発表会」

    2018.11 - 2018.12

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    スーパーサイエンスハイスクール(SSH事業)の一環として行われた高校生による課題研究中間発表会および課題研究発表会において、助言を行った。

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  • 「あさひかわオープンカレッジ」(主催:旭川ウェルビーイング・コンソーシアム事務局)の講師

    2017.11

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    「発がん物質をどのように見つけ出すか ~食品と医薬品の安全性をチェックする~」というテーマで講義を2時間行った。

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Media Coverage

  • ヒト精子 凍結乾燥し保存・再生 旭川医大 水で戻し正常状態に Newspaper, magazine

    北海道新聞  2007.12

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    Author:Other 

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  • 精子保存に「凍結乾燥」 日米大学チーム成功 不妊治療の予備的手法に Newspaper, magazine

    読売新聞(夕刊)  2007.12

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    Author:Other 

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