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UNDIFFERENTIATION PROMOTER AND CRYOPROTECTIVE AGENT USING APROTONIC DIPOLAR ION

Foreign code F210010310
File No. AX02001WO
Posted date 2021年1月29日
Country 世界知的所有権機関(WIPO)
International application number 2020JP018668
International publication number WO 2020230721
Date of international filing 令和2年5月8日(2020.5.8)
Date of international publication 令和2年11月19日(2020.11.19)
Priority data
  • 特願2019-090509 (2019.5.13) JP
Title UNDIFFERENTIATION PROMOTER AND CRYOPROTECTIVE AGENT USING APROTONIC DIPOLAR ION
Abstract 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).
Outline of related art and contending technology 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).
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • Inventor
  • KURODA Kosuke
  • HIRATA Eishu
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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