Production and application of a dental hypoplasia mouse model using a specific organization and a cell disruption system

• Project/Area Number: 23592892
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: Kagoshima University
• Principal Investigator: MATSUMOTO Yuko 鹿児島大学, 医学部・歯学部附属病院, 助教 (20315443)
• Co-Investigator(Kenkyū-buntansha): SATO Masahiro 鹿児島大学, 医用ミニブタ先端開発研究センター, 教授 (30287099) ; SAITOH Issei 新潟大学, 医歯学総合研究科, 准教授 (90404540) ; YAMASAKI Youichi 鹿児島大学, 医歯学総合研究科, 教授 (30200645) ; NOGUCHI Hirofumi 独立行政法人国立病院機構, 千葉東病院臨床研究センター, 研究員 (50378733) ; SAITOH Yoko 新潟大学, 医歯学総合研究科, 助教 (30404487) ; INADA Emi 鹿児島大学, 医学部・歯学部附属病院, 除教 (30448568)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 歯形成 / ameloblast / odontoblast / Tgマウス / odontblast

Research Abstract

We produced a plasmid that specifically expressed a DT-A gene in cells responsible for tooth development, and a construct (pARIC), which connected the red fluorescent gene (RFP) and the Cre gene downstream from amelogenin promoter. In order to check the operation of the plasmid, pARIC and pCETZ-17 were poured into the palate of a two-week-old mouse, and the gene was introduced in vivo by electroporation. The result showed the expression of the lacZ gene indicating that the expression of downstream lacZ gene was the result of the expression of the Cre gene by amelogenin promoter. Moreover, this removed "EGFP cDNA+stopper" inserted by loxP in pCETZ-17.

Research Products

• [Journal Article] The combinational use of CRISPR/Cas9-based gene editing and targeted toxin technology enables efficient biallelic knockout of theα-1,3-galactosyltransferase gene in porcine embryonic fibroblasts (2014)
  • Author(s): Sato M, Miyoshi K, Nagao Y, Nishi Y, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S
  • Journal Title: Xenotransplantation Volume: 21 FEB( Article first published online) Issue: 3 Pages: 291-300
  • DOI: 10.1111/xen.12089
  • Peer Reviewed / Open Access
• [Journal Article] PiggyBac-mediated generation of stable transfectants with surface HLA expression from a small number of cells. (2013)
  • Author(s): Miura H., Inoko H., Inoue I., Okada Y., Tanaka M., Sato M. and Ohtsuka M.
  • Journal Title: Anal. Biochem Volume: 437 Issue: 1 Pages: 29-31
  • DOI: 10.1016/j.ab.2013.02.003
  • Peer Reviewed
• [Journal Article] Mosaic expression of pluripotency-related proteins oct-3/4 and alkaline phosphatase in human pancreatic carcinoma cell PANC-1 (2013)
  • Author(s): Sato M, Maeda S, Inada E, Saitoh I, Kubota N
  • Journal Title: Advanced Studies in Biology Volume: 5 Pages: 157-172
  • URL: http://www.doaj.org/doaj?func=issueTOC&isId=150538&uiLanguage=en
  • Peer Reviewed
• [Journal Article] Hela cells consist of two cell types, as evidenced by cytochemical staining for alkaline phosphatase activity: A possible model for cancer stem cell study. (2013)
  • Author(s): Sato M, Kubota N, Inada E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S
  • Journal Title: Advances in Stem Cells Volume: 2013 Pages: 15-15
  • DOI: 10.5171/2013.208514
  • Peer Reviewed
• [Journal Article] Targeted Toxin-Based Selectable Drug-Free Enrichment of Mammalian Cells with High Transgene Expression (2013)
  • Author(s): Sato M, Akasaka E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S
  • Journal Title: Biology Volume: 2 Issue: 1 Pages: 341-355
  • DOI: 10.3390/biology2010341
  • Peer Reviewed
• [Journal Article] Improvement of hydrodynamics-based gene transfer of nonviral DNA targeted to murine hepatocytes (2013)
  • Author(s): Nakamura S, Maehara T, Watanabe S, Ishihara M, Sato M
  • Journal Title: Biomed Res Int Volume: 2013(2013) Pages: 9-9
  • DOI: 10.1155/2013/928790
  • Peer Reviewed
• [Journal Article] Improvement of pronuclear injection-based targeted transgenesis (PITT) by iCre mRNA-mediated site-specific recombination (2013)
  • Author(s): Ohtsuka M, Miura H, Hayashi H, Nakaoka H, Kimura M, Sato M, Gurumurthy CB, Inoko H
  • Journal Title: Transgenic Research Volume: 22 Issue: 4 Pages: 873-875
  • DOI: 10.1007/s11248-013-9703-x
  • Peer Reviewed
• [Journal Article] Site-targeted non-viral gene delivery by direct DNA injection into the pancreatic parenchyma and subsequent in vivo electroporation in mice. (2013)
  • Author(s): Sato M, Inada E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai T and Watanabe S
  • Journal Title: Biotechnology Journal Volume: 8 Issue: 11 Pages: 1355-1361
  • DOI: 10.1002/biot.201300169
  • Peer Reviewed
• [Journal Article] STO feeder cells are useful for propagation of primarily cultured human deciduous dental pulp cells in view of elimination of contaminated bacteria and promotion of cellular outgrowth. (2013)
  • Author(s): Murakami T, Saitoh I, Inada E, Kurosawa M, Iwase Y, Noguchi H, Terao Y, Yamasaki Y, Hayasaki H, Sato M
  • Journal Title: Cell Medicine Volume: 7 Issue: 1-2 Pages: 75-81
  • DOI: 10.3727/215517913x674234
  • Peer Reviewed
• [Journal Article] Microbial and enzyme technology : an efficient and convenient method for MiniPrep analysis of recombinant plasmids (2013)
  • Author(s): Sato M, Inada E, Saitoh I, Matsumoto Y
  • Journal Title: Journal of Biomedical Science and Engineering Volume: 7 Issue: 03 Pages: 105-107
  • DOI: 10.4236/jbise.2014.73013
  • Peer Reviewed / Open Access
• [Journal Article] Culture conditions for mouse pancreatic stem cells (2013)
  • Author(s): Noguchi H, Saitoh I, Kataoka H, Watanabe M, Noguchi Y, Fujiwara T
  • Journal Title: Cell Medicine Volume: 5 Issue: 2-3 Pages: 63-68
  • DOI: 10.3727/215517913x666495
  • Peer Reviewed
• [Journal Article] Isolation efficiency of mouse pancreatic stem cells is age-dependent (2013)
  • Author(s): Takashi K, Noguchi H, Saitoh I, Kataoka H, Watanabe M, Noguchi Y, Fujiwara T
  • Journal Title: Cell Medicine Volume: 5 Issue: 2-3 Pages: 69-73
  • DOI: 10.3727/215517913x666503
  • Peer Reviewed
• [Journal Article] Comparison of new preservation solution, HN-1, and University of Wisconsin solution in pancreas preservation for porcine islet isolation (2013)
  • Author(s): Akihiro K, Noguchi H, Kuise T, Nakatsuka A, Hirota D, Kataoka H, Kawai T, Inoue K, Imagawa N, Saitoh I, Noguchi Y, Watanabe M, Wada J, Fujiwara T
  • Journal Title: Cell Medicine Volume: 6 Issue: 1-2 Pages: 3-8
  • DOI: 10.3727/215517913x674171
  • Peer Reviewed
• [Journal Article] Maintenance of viability and function of rat islets with the use of ROCK inhibitor Y27632 (2013)
  • Author(s): Kubota Y, Noguchi H, Seita M, Yuasa T, Sasamoto H, Nakaji S, Okitsu T, Fujiwara T, Kobayashi N
  • Journal Title: Cell Medicine Volume: 6 Issue: 1-2 Pages: 15-23
  • DOI: 10.3727/215517913x674199
  • Peer Reviewed
• [Journal Article] Development of Canine Models of Type 1 Diabetes with Partial Pancreatectomy and the Administration of STZ (2013)
  • Author(s): Seita M, Noguchi H, Kubota Y, Kawamoto H, Nakaji S, Kobayashi N, Fujiwara T
  • Journal Title: Cell Medicine Volume: 6 Issue: 1-2 Pages: 25-31
  • DOI: 10.3727/215517913x674289
  • Peer Reviewed
• [Journal Article] Mosaic expression of pluripotency-related proteins oct-3/4 and alkaline phosphatase in human pancreatic carcinoma cell PANC-1. (2013)
  • Author(s): Sato M, Maeda S, Inada E, Saitoh I, Kubota N
  • Journal Title: Advanced Studies in Biology Volume: 5 Pages: 157-172
  • Peer Reviewed
• [Journal Article] Development of a technique for efficient gene transfer to antral follicular cells in the mouse ovary (2012)
  • Author(s): Sato M, Akasaka E, Saitoh I, Ohtsuka M, Watanabe S
  • Journal Title: Systems Biology in Reproductive Medicine Volume: 58 Issue: 3 Pages: 136-141
  • DOI: 10.3109/19396368.2012.656796
  • Peer Reviewed
• [Journal Article] Expression analysis of an α-1,3-galactosyltransferase, an enzyme that creates xenotransplantation-related α-Gal epitope, in pig preimplantation embryos (2012)
  • Author(s): Chi H
  • Journal Title: Animal Science Journal Volume: 83 Issue: 1 Pages: 88-93
  • DOI: 10.1111/j.1740-0929.2011.00964.x
  • Peer Reviewed
• [Journal Article] Successful Suppression of Endogenous <i>&alpha;-1,3-Galactosyltransferase</i> Expression by RNA Interference in Pig Embryos Generated <i>In Vitro</i> (2012)
  • Author(s): Chi H
  • Journal Title: Journal of Reproduction and Development Volume: 58 Issue: 1 Pages: 69-76
  • DOI: 10.1262/jrd.10-165M
  • NAID: 130001170088
  • ISSN: 0916-8818, 1348-4400
  • Peer Reviewed
• [Journal Article] In vivo gene transfer in mouse preimplantation embryos after intraoviductal injection of plasmid DNA and subsequent in vivo electroporation (2012)
  • Author(s): Sato M
  • Journal Title: Syst Biol Reprod Med Volume: 58 Issue: 5 Pages: 278-287
  • DOI: 10.3109/19396368.2012.688088
  • Peer Reviewed
• [Journal Article] Determination of the Optimal Concentration of Several Selective Drugs Useful for Generating Multi-Transgenic Porcine Embryonic Fibroblasts (2012)
  • Author(s): Sato M, Ohtsuka M, Miura H, Miyoshi K, Watanabe S.
  • Journal Title: Reproduction in Domestic Animals/John Wiley & Sons, Inc. Volume: 47 Issue: 5 Pages: 759-765
  • DOI: 10.1111/j.1439-0531.2011.01964.x
  • Peer Reviewed
• [Journal Article] A Cre Knock-in Mouse Line on the Sickle Tail Locus Induces Recombination in the Notochord and Intervertebral Discs. (2012)
  • Author(s): Abe, K. et al.
  • Journal Title: Genesis Volume: 50 Issue: 10 Pages: 758-765
  • DOI: 10.1002/dvg.22035
  • Peer Reviewed
• [Journal Article] CD81 and CD9 work independently as extracellular components upon fusion of sperm and oocyt (2012)
  • Author(s): Ohnami N, Nakamura A, Miyado M, Sato M, Kawano N, Yoshida K, Harada Y, Takezawa Y, Kanai S, Ono C, Takahashi Y, Kimura K, Shida T, Miyado K, Umezawa A
  • Journal Title: Biol Open Volume: 1(7) Issue: 7 Pages: 640-647
  • DOI: 10.1242/bio.20121420
  • Peer Reviewed
• [Journal Article] A simplified protocol for the semi-large scale recovery of plasmids from Escherichia coli grown on agar plates (2012)
  • Author(s): Sato M, Akasaka E, Saitoh I, Ohtsuka M, Nakamura S, Sakurai T, Watanabe S
  • Journal Title: J Biomed Sci Eng. Volume: 5 Issue: 07 Pages: 406-408
  • DOI: 10.4236/jbise.2012.57051
  • Peer Reviewed
• [Journal Article] Generation of a set of mouse STO feeder cell lines that confer resistance to several types of selective drugs (2012)
  • Author(s): Saitoh I, Sato M, Iwase Y, Akasaka E, Yamasaki Y, Noguchi H
  • Journal Title: Cell Medicine Volume: 3(1) Issue: 1-3 Pages: 97-102
  • DOI: 10.3727/215517912x639414
  • Peer Reviewed
• [Journal Article] PITT: Pronuclear Injection-Based Targeted Transgenesis, a Reliable Transgene Expression Method in Mice (2012)
  • Author(s): Ohtsuka M, Miura H, Sato M, Kimura M, Inoko H, Gurumurthy CB
  • Journal Title: Experimental Animals Volume: 61 Issue: 5 Pages: 489-502
  • DOI: 10.1538/expanim.61.489
  • NAID: 10031121853
  • ISSN: 0007-5124, 1341-1357, 1881-7122
  • Peer Reviewed
• [Journal Article] Fluorescent transgenic mice suitable for multi-color aggregation chimera studies (2012)
  • Author(s): Ohtsuka M, Miura H, Gurumurthy CB, Kimura M, Inoko H, Yoshimura S, Sato M
  • Journal Title: Cell and Tissue Research Volume: 350 Issue: 2 Pages: 251-260
  • DOI: 10.1007/s00441-012-1470-0
  • Peer Reviewed
• [Journal Article] Attenuation of limb loss in an experimentally induced hindlimb ischemic model by fibroblast growth factor-2/fragmin/protamine microparticles as a delivery system (2012)
  • Author(s): Nakamura S, Ishihara M, Takikawa M, Kishimoto S, Isoda S, Fujita M, Sato M, Maehara T
  • Journal Title: Tissue Engineering. Part A Volume: 18(21-22) Issue: 21-22 Pages: 2239-2247
  • DOI: 10.1089/ten.tea.2011.0741
  • Peer Reviewed
• [Journal Article] Delivery system for autologous growth factors fabricated with low-molecular-weight heparin and protamine to attenuate ischemic hind-limb loss in a mouse model (2012)
  • Author(s): Nakamura S, Takikawa M, Ishihara M, Nakayama T, Kishimoto S, Isoda S, Ozeki Y, Sato M, Maehara T
  • Journal Title: Journal of Artificial Organs Volume: 15(4) Issue: 4 Pages: 375-385
  • DOI: 10.1007/s10047-012-0658-0
  • Peer Reviewed
• [Journal Article] Targeted transgenesis through pronuclear injection of improved vectors into in vitro fertilized eggs (2011)
  • Author(s): Ohtsuka, M., Miura, H., Nakaoka, H., Kimura, M., Sato, M., Inoko, H.
  • Journal Title: Transgenic Research Volume: 21:1 Issue: 1 Pages: 225-226
  • DOI: 10.1007/s11248-011-9505-y
  • Peer Reviewed
• [Journal Article] Condyle Shape in Relation to Anterior Disk Displacement in young adolescence (2011)
  • Author(s): Hiroko Hasegawa, Issei Saitoh, Kuniko Nakakura-Ohshima, Koki Shigeta Toshihiro Yoshihara, Shigeaki Suenaga, Yuko Matsumoto, Emi Inada, Youichi Yamasaki
  • Journal Title: Journal of Craniomandibular Practice Volume: 29 Pages: 100-110
  • Peer Reviewed
• [Journal Article] Liver lobe- and strain-difference in gene expression after hydrodynamics-based gene delivery in mice
  • Author(s): Nakamura S, Maehara T, Ishihara M, Sato M
  • Journal Title: Animal Biotechnology Volume: (in press)
  • Peer Reviewed
• [Presentation] Multipotency of juvenile human buccal epithelial cells (2014)
  • Author(s): Murakami T, Saitoh I, Iwase Y, Inada E, Mastuyama J, Ohshima H, Hayasaki H, Sato M
  • Organizer: AADR
  • Place of Presentation: North Carolina
• [Presentation] Multipotency of juvenile human buccal epithelial cells. (2014)
  • Author(s): T Murakami, I Saitoh, Y Iwase, E Inada, J Mastuyama, H Ohshima, H Hayasaki M Sato
  • Organizer: 43rd Annual Meeting & Exibition of the AADR (American Association for Dental Research)
  • Place of Presentation: America (North Carolina)
• [Presentation] gcr2タンパクは糖鎖を介してI型BMP受容体と結合しシグナル伝達制御に関与する (2013)
  • Author(s): 渡部聡、梶原景正、麥倉真一郎、桜井敬之、中村伸吾、木村穣、佐藤正宏
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] RISPR/Cas9による遺伝子編集と標的毒素法との組み合わせは、α-1,3-galactosyltransferase遺伝子を完全にKOしたブタ胎仔性線維芽細胞の効率的作製に有効である (2013)
  • Author(s): 佐藤正宏、三好和睦、長尾洋三、西洋平、大塚正人、中村伸吾、桜井敬之、渡部聡
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] Syngenic grafting of a whole male juvenile gonadal tissue into the adult testes confers successful spermatogenesis in mice (2013)
  • Author(s): Tada N, Kanai F, Nakamura E, Lu H, Saito M, Sato M
  • Organizer: The 3rd World Congress of the International Society for Fertility Preservation
  • Place of Presentation: Malia Valencia Hotel, Valencia, Spain
• [Presentation] トランスポゾンPiggyBacシステムによる複数遺伝子のブタ細胞への同時導入 (2013)
  • Author(s): 郡山美優, 稲田絵美, 齋藤一誠, 三浦浩美, 大塚正人, 中村伸吾, 桜井敬之, 渡部聡, 三好和睦, 佐藤正宏
  • Organizer: 第106回日本繁殖生物学会
  • Place of Presentation: 東京
• [Presentation] ヒト乳歯歯髄由来iPS細胞樹立におけるフィーダー細胞選択の重要性 (2013)
  • Author(s): 村上智哉, 齊藤一誠, 稲田絵美, 岩瀬陽子, 長谷川大子, 窪田直子, 松本祐子, 大島邦子,岡暁子, 山崎要一, 早崎治明
  • Organizer: 第51回日本小児歯科学会
  • Place of Presentation: 岐阜
• [Presentation] 受精卵へのiCre mRNA注入によるターゲットトランスジェネシス法(PITT法)の効率改善 (2013)
  • Author(s): 大塚正人、三浦浩美、佐藤正宏、木村穣
  • Organizer: 第60回日本実験動物学会総会
  • Place of Presentation: 茨城
• [Presentation] ヒト乳歯歯髄由来iPS細胞樹立におけるフィーダー細胞選択の重要性 (2013)
  • Author(s): 村上智哉, 齊藤一誠, 稲田絵美, 岩瀬陽子, 長谷川大子, 窪田直子, 松本祐子, 大島邦子, 岡 暁子, 山崎要一, 早崎治明
  • Organizer: 第５１回日本小児歯科学会
  • Place of Presentation: 岐阜県
• [Presentation] 初代ヒト乳歯歯髄細胞における幹細胞特異的遺伝子発現の探索 (2012)
  • Author(s): 乃村俊樹, 齊藤一誠, 稲田絵美, 長谷川大子, 松本祐子, 窪田直子, 山﨑要一
  • Organizer: 第30回日本小児歯科学会九州地方会大会
  • Place of Presentation: 長崎