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PRODUCTION METHOD AND KIT OF INDUCED PLURIPOTENT STEM CELLS

Foreign code F210010283
File No. (S2019-0335-N0)
Posted date Jan 28, 2021
Country WIPO
International application number 2020JP016924
International publication number WO 2020213725
Date of international filing Apr 17, 2020
Date of international publication Oct 22, 2020
Priority data
  • P2019-078907 (Apr 17, 2019) JP
Title PRODUCTION METHOD AND KIT OF INDUCED PLURIPOTENT STEM CELLS
Abstract This method of producing induced pluripotent stem cells involves a step for introducing an initialization factor into somatic cells of a mammal, and culturing in a neural stem cell culture medium to obtain induced neural stem cell-like cells, and a step for cultivating said induced neural stem cell-like cells in a growth medium to obtain induced pluripotent stem cells , wherein the initialization factor contains an OCT family, a SOX family, a KLF family, a MYC family, a LIN28 family and a P53 function inhibitor.
Outline of related art and contending technology BACKGROUND ART
Techniques for establishing iPS cells from human and mouse somatic cells have been improved and simplified, and the efficiency of establishing iPS cells has been improved. On the other hand, there is an animal (pluripotent induction resistant animal) which is difficult to establish iPS cells from adult somatic cells. Dogs, pigs, marmosets, cats, cows, horses, and the like are known as pluripotent induction resistant animals.
See), for example, (, Non-Patent Document 1, in which canine iPS cells can be established by persistently expressing a reprogramming factor in canine fetus fibroblasts. In addition, see), for example, Non-Patent Document 2, it is possible to establish canine iPS cells from canine fetus fibroblasts by using Sendai virus. See), for example, Non-Patent Document 3, in which porcine iPS cells can be established by persistently expressing a reprogramming factor in porcine fetus fibroblasts.
Scope of claims (In Japanese)[請求項1]
 哺乳動物の体細胞に初期化因子を導入し、神経幹細胞培養培地で培養して誘導神経幹細胞様細胞を得る工程と、
 前記誘導神経幹細胞様細胞を増殖用培地で培養して誘導多能性幹細胞を得る工程と、を含み、
 前記初期化因子が、OCTファミリー、SOXファミリー、KLFファミリー、MYCファミリー、LIN28ファミリー及びP53機能阻害物質を含む、誘導多能性幹細胞の製造方法。

[請求項2]
 前記初期化因子が、更に、KDMファミリー、GLISファミリー及びNANOGからなる群より選択される一種以上を含む、請求項1に記載の誘導多能性幹細胞の製造方法。

[請求項3]
 前記体細胞が成体由来である、請求項1又は2に記載の製造方法。

[請求項4]
 前記神経幹細胞培養培地が、MAPキナーゼ阻害剤、GSK3β阻害剤及びTGFβ阻害剤を含む、請求項1~3のいずれか一項に記載の製造方法。

[請求項5]
 前記増殖用培地が、bFGFを含む、請求項1~4のいずれか一項に記載の製造方法。

[請求項6]
 請求項1~5のいずれか一項に記載の製造方法により製造された、誘導多能性幹細胞。

[請求項7]
 OCTファミリー、SOXファミリー、KLFファミリー、MYCファミリー、LIN28ファミリー及びP53機能阻害物質を含む初期化因子と、
 MAPキナーゼ阻害剤、GSK3β阻害剤及びTGFβ阻害剤を含む神経幹細胞培養培地と、を含む、
 多能性誘導耐性動物の体細胞から誘導多能性幹細胞を製造するためのキット。

[請求項8]
 前記初期化因子が、更に、KDMファミリー、GLISファミリー及びNANOGからなる群より選択される一種以上を含む、請求項7に記載のキット。

[請求項9]
 bFGFを含む増殖用培地を更に含む、請求項7又は8に記載のキット。
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • KEIO UNIVERSITY
  • NIHON UNIVERSITY
  • Inventor
  • OKANO Hideyuki
  • SHIOZAWA Seiji
  • YOSHIMATSU Sho
  • EDAMURA Kazuya
  • IGUCHI Aozora
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN WS ZA ZM ZW
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
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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