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1. WO2020179380 - PREPARATION AND EXPANSION CULTURE OF ENDOTHELIAL PROGENITOR CELL

Publication Number WO/2020/179380
Publication Date 10.09.2020
International Application No. PCT/JP2020/005255
International Filing Date 12.02.2020
Chapter 2 Demand Filed 26.05.2020
IPC
A61P 9/00 2006.01
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
9Drugs for disorders of the cardiovascular system
C12N 5/071 2010.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
07Animal cells or tissues
071Vertebrate cells or tissues, e.g. human cells or tissues
C12N 5/0735 2010.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
07Animal cells or tissues
071Vertebrate cells or tissues, e.g. human cells or tissues
073Embryonic cells or tissues; Foetal cells or tissues
0735Embryonic stem cells; Embryonic germ cells
C12N 5/10 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
10Cells modified by introduction of foreign genetic material, e.g. virus-transformed cells
A61K 35/44 2015.01
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
35Medicinal preparations containing materials or reaction products thereof with undetermined constitution
12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
44Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
Applicants
  • 公立大学法人名古屋市立大学 PUBLIC UNIVERSITY CORPORATION NAGOYA CITY UNIVERSITY [JP]/[JP]
Inventors
  • 松永 民秀 MATSUNAGA Tamihide
  • 坡下 真大 HASHITA Tadahiro
  • 青木 啓将 AOKI Hiromasa
Agents
  • 萩野 幹治 HAGINO Mikiharu
Priority Data
2019-04011706.03.2019JP
Publication Language Japanese (JA)
Filing Language Japanese (JA)
Designated States
Title
(EN) PREPARATION AND EXPANSION CULTURE OF ENDOTHELIAL PROGENITOR CELL
(FR) PROCÉDÉ DE PRÉPARATION ET DE CULTURE À GRANDE ÉCHELLE DE CELLULES PROGÉNITRICES D'ENDOTHÉLIUM VASCULAIRE
(JA) 血管内皮前駆細胞の調製及び拡大培養
Abstract
(EN)
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.
(FR)
L'invention a pour objet de fournir un moyen de préparer des cellules progénitrices d'endothélium vasculaire d'une grande pureté de manière simple et à bas coût. En outre, l'invention a pour objet de fournir un procédé de prolifération efficace des cellules progénitrices d'endothélium vasculaire. Selon l'invention, les cellules progénitrices d'endothélium vasculaire d'une grande pureté sont préparées via : une étape au cours de laquelle des cellules souches pluripotentes sont différenciées en cellules progénitrices d'endothélium vasculaire ; et une étape au cours de laquelle les cellules progénitrices d'endothélium vasculaire sont purifiées en mettant en œuvre la différence de capacité d'adhésion des cellules progénitrices d'endothélium vasculaire configurant la population cellulaire obtenue à l'étape précédente et des autres cellules. En outre, les cellules progénitrices d'endothélium vasculaire sont ajoutées à un facteur de prolifération de fibroblaste basique et à un facteur de croissance épithélial, et sont cultivées et prolifèrent en présence d'un inhibiteur ROCK, d'un inhibiteur GSK-3β et d'un inhibiteur de récepteur TGF-β.
(JA)
簡便且つ低コストで高純度の血管内皮前駆細胞を調製する手段を提供することを課題とする。また、血管内皮前駆細胞を効率的に増殖させる方法を提供することも課題とする。多能性幹細胞を血管内皮前駆細胞へと分化させる工程と、該工程で得られた細胞集団を構成する血管内皮前駆細胞とその他の細胞の接着能力の差を利用して血管内皮前駆細胞を純化する工程によって、高純度の血管内皮前駆細胞を調製する。一方、血管内皮前駆細胞を、塩基性線維芽細胞増殖因子と上皮成長因子に加え、ROCK阻害剤、GSK-3β阻害剤及びTGF-β受容体阻害剤の存在下で培養し、増殖させる。
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