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1. WO2021020720 - EQUINE AMNIOTIC FLUID-DERIVED STEM CELLS, AND CELL THERAPEUTIC AGENT FOR REGENERATING MUSCULOSKELETAL SYSTEM, COMPRISING SAME

Publication Number WO/2021/020720
Publication Date 04.02.2021
International Application No. PCT/KR2020/007476
International Filing Date 10.06.2020
IPC
C12N 5/0775 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
077Mesenchymal cells, e.g. bone cells, cartilage cells, marrow stromal cells, fat cells or muscle cells
0775Mesenchymal stem cells; Adipose-tissue derived stem cells
A61K 35/28 2006.01
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
28Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
A61P 21/00 2006.01
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
21Drugs for disorders of the muscular or neuromuscular system
CPC
A61K 35/28
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
28Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
A61P 21/00
AHUMAN NECESSITIES
61MEDICAL OR VETERINARY SCIENCE; HYGIENE
PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
21Drugs for disorders of the muscular or neuromuscular system
C12N 2533/54
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
2533Supports or coatings for cell culture, characterised by material
50Proteins
54Collagen; Gelatin
C12N 5/0668
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;
06Animal cells or tissues; Human cells or tissues
0602Vertebrate cells
0652Cells of skeletal and connective tissues; Mesenchyme
0662Stem cells
0668Mesenchymal stem cells from other natural sources
Applicants
  • 주식회사 엠케이바이오텍 MKBIOTECH CO., LTD. [KR]/[KR]
  • 충남대학교 산학협력단 THE INDUSTRY & ACADEMIC COOPERATION IN CHUNGNAM NATIONAL UNIVERSITY (IAC) [KR]/[KR]
Inventors
  • 김민규 KIM, Minkyu
  • 김은영 KIM, Eunyeong
  • 이은지 LEE, Eunji
  • 김동언 KIM, Dongeon
  • 남윤석 NAM, Yoonseok
  • 길태영 GIL, Taeyoung
Agents
  • 길준연 GIL, Jun Yeoun
Priority Data
10-2019-009119326.07.2019KR
Publication Language Korean (KO)
Filing Language Korean (KO)
Designated States
Title
(EN) EQUINE AMNIOTIC FLUID-DERIVED STEM CELLS, AND CELL THERAPEUTIC AGENT FOR REGENERATING MUSCULOSKELETAL SYSTEM, COMPRISING SAME
(FR) CELLULES SOUCHES DÉRIVÉES DE LIQUIDE AMNIOTIQUE ÉQUIN, ET AGENT THÉRAPEUTIQUE CELLULAIRE POUR RÉGÉNÉRER LE SYSTÈME MUSCULO-SQUELETTIQUE LES COMPRENANT
(KO) 말과 동물 양수 유래 줄기세포 및 이를 함유하는 근골격계 재생용 세포 치료제
Abstract
(EN)
The present invention relates to equine fetal tissue-derived stem cells, and a cell therapeutic agent for regenerating a musculoskeletal system, comprising same, and, more particularly, to: a method for preparing equine amniotic fluid-derived stem cells; the analysis of characteristics of amniotic fluid stem cells prepared by the method; equine amniotic fluid-derived stem cells comprising the stem cells as an active ingredient; and a cell therapeutic agent for regenerating a musculoskeletal system, comprising same. The present invention provides equine amniotic fluid-derived stem cells. In addition, the present invention provides a method for preparing equine amniotic fluid-derived mesenchymal stem cells, comprising the steps of: (step 1) collecting amniotic fluid from a pregnant horse; (step 2) isolating monocytes from the collected amniotic fluid; and (step 3) initially culturing the isolated amniotic fluid-derived monocytes. Moreover, the present invention provides a method for preparing equine mesenchymal stem cells, comprising the steps of: (step 1) collecting amniotic fluid from a pregnant horse; (step 2) isolating monocytes from the collected amniotic fluid; (step 3) initially culturing the isolated amniotic fluid-derived monocytes; (step 4) culturing the initially cultured equine amniotic fluid-derived cells of passages 3 to 5 by using the culture medium Dulbecco's Modified Eagle Medium (DMEM), which contains 10-20% fetal bovine serum (FBS), 0.5-1.5% glutamax, and 0.5-1.5% penicillin/streptomycin; and (step 5) removing the medium in a culture dish, washing same with phosphate buffered saline (PBS, pH 7.2), and then separating cells from the basal surface by using 0.01-0.1% Typsin-EDTA. The present invention also provides equine amniotic fluid-derived stem cells having a musculoskeletal system treatment function. Furthermore, the present invention provides a cell therapeutic agent for regenerating a musculoskeletal system, comprising the equine amniotic fluid-derived stem cells.
(FR)
La présente invention concerne des cellules souches issues d'un tissu foetal équin, et un agent thérapeutique cellulaire pour la régénération d'un système musculo-squelettique les comprenant, et plus particulièrement, un procédé de préparation de cellules souches dérivées du liquide amniotique équin; l'analyse de caractéristiques de cellules souches de liquide amniotique préparées par le procédé; des cellules souches dérivées de liquide amniotique équin comprenant les cellules souches en tant que principe actif; et un agent thérapeutique cellulaire pour régénérer un système musculo-squelettique comprenant lesdites cellules. La présente invention concerne des cellules souches dérivées du liquide amniotique équin. De plus, la présente invention concerne un procédé de préparation de cellules souches mésenchymateuses dérivées d'un liquide amniotique équin, comprenant les étapes suivantes: (Étape 1) collecte du liquide amniotique provenant d'une jument enceinte; (étape 2) isolement des monocytes du liquide amniotique collecté; et (étape 3) culture dans un premier temps des monocytes dérivés de liquide amniotique isolés. De plus, la présente invention concerne un procédé de préparation de cellules souches mésenchymateuses équines, comprenant les étapes consistant à : (Étape 1) collecte du liquide amniotique provenant d'une jument enceinte; (étape 2) isolement des monocytes du liquide amniotique collecté; (étape 3) culture dans un premier temps des monocytes dérivés de liquide amniotique isolés; (étape 4) culture des cellules issues du liquide amniotique équin, initialement cultivées, de passages 3 à 5 en utilisant le milieu de culture Dulbecco's Modified Eagle (DMEM), contienant de 10 à 20 % de sérum bovin foetal (FBS), 0,5 à 1,5 % de glutamax, et 0,5 à 1,5 % de pénicilline/streptomycine; et (étape 5) élimination du milieu dans un récipient de culture, son lavage avec une solution saline tamponnée au phosphate (PBS, pH 7,2), puis la séparation des cellules de la surface de base à l'aide de 0,01 à 0,1 % de Typsin-EDTA. La présente invention concerne également des cellules souches dérivées du liquide amniotique équin ayant une fonction de traitement du système musculo-squelettique. En outre, la présente invention concerne un agent thérapeutique cellulaire pour la régénération d'un système musculo-squelettique, comprenant les cellules souches dérivées du liquide amniotique équin.
(KO)
본 발명은 말과 동물의 태아 조직 유래 줄기세포(Equine fetal tissue derived stem cells)및 이를 함유하는 근골격계 재생용 세포치료제에 관한 것으로, 보다 구체적으로는, 말과 동물의 양수 유래 줄기세포(Equine amniotic fluid stem cells)의 제조방법, 상기 방법으로 제조된 양수 줄기세포의 특성 분석, 상기 줄기세포를 유효 성분으로 함유하는 말 양수 유래 줄기세포 및 이를 함유하는 근골격계 재생용 세포 치료제에 관한 것이다. 본 발명은 말과 동물 양수 유래의 줄기세포를 제공한다. 또한 본 발명은 임신한 말의 양수를 채취하는 과정(1과정), 상기의 채취한 양수에서 단핵 세포를 분리하는 과정(2과정), 상기의 분리된 양수 유래 단핵세포를 초기 배양하는 과정(3과정)을 포함하는 말과 동물 양수 유래의 중간엽 줄기세포를 제조하는 방법를 제공한다. 또한 본 발명은 임신한 말의 양수를 채취하는 과정(1과정), 상기의 채취한 양수에서 단핵 세포를 분리하는 과정(2과정), 상기의 분리된 양수 유래 단핵세포를 초기 배양하는 과정(3과정), 상기에서 초기 배양된 3~5계대 사이의 말 양수 유래 세포를 10~20 % FBS (Fetal Bovine Serum), 0.5~1.5 % Glutamax 및 0.5~1.5 % 페니실린/스트렙토마이신이 포함된 Dulbecco Modified Eagle Medium(DMEM) 배양 배지를 사용하여 배양하는 과정(4과정), 상기의 배양접시 내의 배지를 제거하고 PBS (Phosphate buffered saline, PH 7.2)으로 세척 후 0.01~0.1% Typsin-EDTA를 이용하여 세포를 기저면에서 분리시키는 과정(5과정)을 포함한 말과 동물 중간엽 줄기세포를 제조하는 방법을 제공한다. 또한 본 발명은 근골격계 치료 기능이 있는 것을 특징으로 하는 말과 동물 양수 유래의 줄기세포를 제공한다. 또한 본 발명은 상기한 말과 동물 양수 유래의 줄기세포를 포함하는 근골격계 재생용 세포 치료제를 제공한다.
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