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METHOD FOR PRODUCING CANINE iPS CELLS 新技術説明会

外国特許コード F170008953
整理番号 (S2015-1659-N0)
掲載日 2017年2月16日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP068194
国際公開番号 WO 2016204298
国際出願日 平成28年6月19日(2016.6.19)
国際公開日 平成28年12月22日(2016.12.22)
優先権データ
  • 特願2015-123775 (2015.6.19) JP
発明の名称 (英語) METHOD FOR PRODUCING CANINE iPS CELLS 新技術説明会
発明の概要(英語) [Problem] To produce, by using a liquid culture having an extremely simple composition, a canine iPS cell line that can be subcultured stably for a long period of time.
[Solution] The present invention obtains canine iPS cells by culturing canine somatic cells having introduced therein four transcription factors of OCT3/4, SOX2, KLF4, and C-MYC that are nuclear reprogramming factors by means of a drug-inducible vector that can control expression of the transcription factors through usage of, for example, doxycycline, in a liquid culture containing basic fibroblast growth factor (bFGF) but not containing a pluripotency maintaining factor (not including basic fibroblast growth factor) such as LIF, and further preferably not containing a differentiation suppressor such as MEK suppressor and GSK3β suppressor.
従来技術、競合技術の概要(英語) BACKGROUND ART
Dogs, rodent such as a mouse or a rat compared to close to physiological conditions in addition to the human, large size, service life is also long, can be administered in a relatively large amount of a drug, is also suited to a long term experiment. In addition, living in the human and in the same environment and the human disease model animals as it is more . Further, canine genetic diseases found in more than half of the same gene mutation similar to human disease, canine genomic sequence was elucidated from the fact that, as a disease model for the demand of the dogs may be further extended.
So far, the fabrication of canine iPS cells but various report as an example, in these reports, a basal medium Ham's F-12 DMEM/F12(Dulbecco's Modified Eagle medium medium were mixed with each other by equal amount) to, such as KSR of serum such as FBS or serum replacement, further LIF on the differentiation of a reducing agent such as have been used as the culture medium. For example, in Patent Document 1, basal MAPKK (mitogen-activated protein kinase kinase) inhibitor and PD0325901 inhibitor ALK (activin receptor-like kinase) and A-83-01 GSK (glycogen synthase kinase) inhibitor HDAC and LIF (leukemia inhibitory factor) and CHIR99021 (histone deacetylase) inhibitor valproic acid mixed with a culture medium is used. In addition, in Patent Document 2, basal medium and serum substitute KSR LIF and bFGF (basic fibroblast growth factor) a mixture of a culture medium is used. Non-patent document 1 is further, and basal bFGF and LIF KSR, further GSK3 β (glycogen, synthase, kinase 3 β) inhibitor, MEK (ERK-MAP kinase pathway blockers) suppressor, TGF-β antagonist (beta growth factor mutant form) a mixture of a culture medium is used.
On the other hand, in human iPS cells and mouse iPS cells, without using bFGF, neural cell culture medium is a basal DMEM/F12 supplement N2 (trade name) used in a mixture of neurobasal medium and B27 medium supplement (trade name) of the culture media mixture to mixing LIF, MEK inhibitor, medium GSK3 β inhibitor used (non-patent document 2, 3). In these documents, the resulting iPS colonies to an enzyme treatment can be a single-cell can be obtained, the single culture of the cells can be about 50 and margin is.
Further, in non-patent document 4 and 5, undifferentiated mouse iPS cells than similar to having a three-dimensional colony forming ability when it is reported that, by enzymatic passaging method cannot be resistant to, if the amount of each inhibitor was added to the medium cannot be maintained its form. In addition, a long period of time may not be from the culture, the strains of these iPS cells has not been considered.
To date, such as LIF or maintain pluripotent differentiation SK3 β inhibitors such as inhibitors on the differentiation factors MEK without using factors, cell growth-promoting factor such as bFGF in the presence of only, dogs from body cells to produce iPS cells, iPS cells can be subcultured over a long period has been no report of produced.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • OSAKA PREFECTURE UNIVERSITY PUBLIC CORPORATION
  • 発明者(英語)
  • INABA Toshio
  • NISHIMURA Toshiya
  • TANAKA Erina
  • HATOYA Shingo
  • SUGIURA Kikuya
国際特許分類(IPC)
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ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
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