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UNDIFFERENTIATION PROMOTER AND CRYOPROTECTIVE AGENT USING APROTONIC DIPOLAR ION

外国特許コード F210010310
整理番号 AX02001WO
掲載日 2021年1月29日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2020JP018668
国際公開番号 WO 2020230721
国際出願日 令和2年5月8日(2020.5.8)
国際公開日 令和2年11月19日(2020.11.19)
優先権データ
  • 特願2019-090509 (2019.5.13) JP
発明の名称 (英語) UNDIFFERENTIATION PROMOTER AND CRYOPROTECTIVE AGENT USING APROTONIC DIPOLAR ION
発明の概要(英語) The purpose of the present invention is to provide an undifferentiation promoter that dissolves a chemical agent in a manner similar to DMSO and the like, that does not induce differentiation of cells when being added to a medium, and that is capable of promoting undifferentiation. This undifferentiation promoter is characterized by containing an aprotonic dipolar ion represented by formula (1) (in the formula, A represents an anion selected from the group consisting of SO3-, -COO-, -OP=O(H)O-, -OP=O(CH3)O-, and -OP=O(OR3)O-, R1 represents an alkyl group having 1-8 carbon atoms and optionally including 1 or 2 oxygen atoms in the molecule chain, R2 represents an alkylene group having 3-5 carbon atoms, and R3 represents hydrogen or an alkyl group optionally having a heteroatom in the molecule chain).
従来技術、競合技術の概要(英語) BACKGROUND ART
Conventionally, when adding a hydrophobic agent to a culture medium in an assay using cells, the hydrophobic agent is dissolved in a solvent such as dimethyl sulfoxide (DMSO) and then dispersed in the culture medium. However, DMSO is known to induce differentiation of stem cells (NpL 1 ~ 3). In contrast, "Cellartis (®) DEF-CS 500 Culture System" available from Takara Bio Inc. and the like are provided as a culture medium for maintaining an undifferentiated state, but there is room for further improvement.
DMSO and glycerol are also widely used as cryoprotectants when cryopreserving cells, and have been shown to be the most effective reagents for protecting cells and organelles. The cryoprotectant suppresses the growth of ice crystals formed in cells when the cells are frozen. DMSO and the like are cell-permeable and retard the rate of growth of ice crystals in and out of the cell and inhibit ice crystal formation. DMSO and glycerol, however, are physiologically toxic and are either infused with cells to a recipient or are known to cause high blood pressure, vomiting and vomiting due to the handling of the personnel.
Furthermore, in order to increase the viability of the cells, a cryoprotectant such as DMSO is often further mixed with a protein such as bovine serum albumin. However, serum is at risk of being contaminated with viruses or the like, and biological activity varies depending on lot, so (Patent Document 1), which takes significant effort to select excellent lots. Furthermore, there is concern about rejection in regenerative medicine by using a protein derived from a different animal.
Meanwhile, cryopreservation of sperm is considered an important problem in livestock industry, but it is proposed to use betaine or carnitine in place of a cryopreservative such as glycerin in order to maintain the mobility of sperm, but proteins derived from different animals such as egg yolk are used in the culture medium in both (Patent Documents 2 and 3).
  • 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
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • KURODA Kosuke
  • HIRATA Eishu
国際特許分類(IPC)
指定国 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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