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PREPARATION AND EXPANSION CULTURE OF ENDOTHELIAL PROGENITOR CELL

Foreign code F200010250
File No. (S2019-0238-N0)
Posted date 2020年10月30日
Country 世界知的所有権機関(WIPO)
International application number 2020JP005255
International publication number WO2020179380
Date of international filing 令和2年2月12日(2020.2.12)
Date of international publication 令和2年9月10日(2020.9.10)
Priority data
  • 特願2019-040117 (2019.3.6) JP
Title PREPARATION AND EXPANSION CULTURE OF ENDOTHELIAL PROGENITOR CELL
Abstract The present invention addresses the problem of providing a means for easily preparing endothelial progenitor cells at high purity and low cost. The present invention also addresses the problem of providing a method for efficiently proliferating endothelial progenitor cells. Endothelial progenitor cells are prepared at high purity by the steps of: differentiating pluripotent stem cells into endothelial progenitor cells; and purifying the endothelial progenitor cells using the difference in adhesion ability between the endothelial progenitor cells of the cell population obtained in the previous step and other cells. Meanwhile, the endothelial progenitor cells are cultured and proliferated in the presence of a ROCK inhibitor, a GSK-3β inhibitor, and a TGF-β receptor inhibitor as well as a basic fibroblast growth factor and an epidermal growth factor.
Outline of related art and contending technology BACKGROUND ART
Human pluripotent stem cells including induced pluripotent stem cells (induced pluripotent stem cells , iPS cells) and embryonic stem cells (embryonic stem cells , ES cells) proliferate infinitely and have the ability to differentiate into any site of the human body. Because of this characteristic, human pluripotent stem cells (, particularly iPS cells), are expected to be used for the construction of drug screening models simulating human organs and clinical applications in regenerative medicine.
Various clinical applications of the iPS cell-derived vascular endothelial progenitor cells (EPCs) may be considered, such as blood vessel regeneration, myocardial infarction treatment by co-transplantation with myocardium, and three-dimensional cellular tissue construction of advanced organs. In addition, it is expected to construct a pathological model of vascular diseases and to use it as a screening system in developing new drugs by differentiating it into brain capillary endothelial cells. A large amount of high-purity vascular endothelial progenitor cells are required to be used for such purposes.
Reference) is now made to (, for example, Non-Patent Documents 1 and 2, where many reports have been made that vascular endothelial cells (ECs) and vascular endothelial progenitor cells are efficiently differentiated and induced from ES and iPS cells. However, since the rate of differentiation into vascular endothelial cells and vascular endothelial progenitor cells is greatly affected by the cell line used and the state before differentiation, it is often purified by using antibody beads/magnetic beads or cell sorter in order to obtain high-purity cells. In addition, efficient expansion culture of iPS cell-derived vascular endothelial progenitor cells has been reported. The research group of the present inventors reported a novel method for inducing differentiation of vascular endothelial progenitor cells by applying iPS-sac method in the previous patent application (Patent Document 1). Patent Document 2 discloses a method for preparing vascular endothelial progenitor cells from embryonic stem cells.
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • NAGOYA CITY UNIVERSITY
  • Inventor
  • MATSUNAGA Tamihide
  • HASHITA Tadahiro
  • AOKI Hiromasa
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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