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METHOD FOR REESTABLISHING PLURIPOTENT STEM CELLS

Foreign code F160008805
File No. (S2014-1604-N0)
Posted date Aug 4, 2016
Country WIPO
International application number 2015JP078699
International publication number WO 2016052759
Date of international filing Oct 2, 2015
Date of international publication Apr 7, 2016
Priority data
  • P2014-203679 (Oct 2, 2014) JP
Title METHOD FOR REESTABLISHING PLURIPOTENT STEM CELLS
Abstract The present application pertains to a method for reestablishing stem cells having a chimera-forming capacity, and to cells obtained by said method. This method is a technique for improving stem cell quality by monoclonally replicating stem cells having a chimera-forming capacity that are selected from a heterogeneous cell population.
Scope of claims [claim1]
1.  A method of re-establishing the stem cells with a chimeric forming ability between different, the following steps:  (I) the first mammalian species derived from pluripotent (pluripotent) stem cells or multipotent properties (multipotent) stem cells, co-cultured with high-quality pluripotent stem cells derived from the seeds of the second mammal;  In co-cultures of (ii) Step (i), to form the cell aggregates, and sorting the cell group containing stem cells derived from pluripotent stem cells or multipotent stem cells from the species of the first mammal to;  (Iii) co-cultured with host embryo of a group of cells derived from the seeds of the second mammalian step (ii);  (Iv) separating the inner cell mass from the step (iii) co-cultured host embryo; and  (V) by cloning the stem cells derived from the inner cell mass in pluripotent stem cells or multipotent stem cells derived from the seeds of the first mammal, to re-establish the stem cells with a chimeric forming ability between heterogeneous; Includes, here in the first mammalian species and the second mammalian species is a different species, and the pluripotent stem cells or multipotent stem cells derived from a first mammalian species, rodent except if it is a pluripotent stem cells or multipotent stem cells derived from mammalian species, it is a rodent-derived induced pluripotent stem cells (iPS cells) or multipotent stem cells, the method.
[2]
[claim2]
2.  Further comprises re-established stem cells in step (v), to give a cell population combined with high-quality pluripotent stem cells from species of the second mammal, repeating steps (iii) ~ (v) the method of claim 1.
[3]
[claim3]
3.  Pluripotent stem cells, the following: is selected from the group consisting of ES cells and induced pluripotent stem cells (iPS cells), and multipotent stem cells following: vegetative Memiki cells (TS cells), the epiblast stem cells (Epis cells ), embryonic germ cells (EG cells), pluripotent germ cells (mGS cells) cloned embryo-derived ES cells (ntES cells), hematopoietic stem cells, neural stem cells, and mesenchymal stem cells are selected from the group consisting of that method according to claim 1 or 2.
[4]
[claim4]
4.  Pluripotent stem cells or multipotent stem cells from species of the first mammal is an ES cell or iPS cells, the method according to any one of claims 1 to 3.
[5]
[claim5]
5.  High quality pluripotent stem cells are naive type of pluripotent stem cells, the method according to any one of claims 1 to 4.
[6]
[claim6]
6.  Host embryo, early embryos, tetraploid embryos, male embryos, parthenogenetic embryo and placenta contributing ES cells are selected from the group consisting of A method according to any one of claims 1 to 5 .
[7]
[claim7]
7.  Step (iii) is microinjected or aggregation of cell group to a second early embryos or tetraploid embryos from mammalian species step (ii) is performed by co-culturing with any of claims 1 to 6 method according to one of claims 1.
[8]
[claim8]
8.  The first mammalian species and a second mammalian species, respectively, mice, rats, rabbits, dogs, cats, horses, cows, goats, sheep, pigs, monkeys, selected from the group consisting of human, claim 1 to the method of any one of 7.
[9]
[claim9]
9.  A method of re-establishing the stem cells with a chimeric forming ability between different, the following steps:  (I) a first mammalian species derived multipotent (pluripotent) stem cells or multipotent properties (multipotent) stem cells, obtaining a cell population combined with high-quality pluripotent stem cells from species of the second mammalian ;  (Ii) a cell population of step (i), co-cultured with host embryo from species of the second mammal;  (Iii) separating the inner cell mass from the step (ii) co-cultured host embryo; and  By cloning the stem cells derived from (iv) the inner cell mass in pluripotent stem cells or multipotent stem cells derived from the seeds of the first mammal, to re-establish the stem cells with a chimeric forming ability between heterogeneous; Includes, here in the first mammalian species and the second mammalian species is a different species, and the pluripotent stem cells or multipotent stem cells derived from a first mammalian species, rodent except if it is a pluripotent stem cells or multipotent stem cells derived from mammalian species, it is a rodent-derived induced pluripotent stem cells (iPS cells) or multipotent stem cells, the method.
[10]
[claim10]
10.  Comprising repeating the steps (i) ~ (iv), this time, the second and subsequent cycles step (i), the stem cells were re-established in step (iv) of the immediately preceding cycle, the second mammalian species performed by obtaining a cell group combined with high-quality-derived multipotent stem cells, the method of claim 9.
[11]
[claim11]
11.  First of pluripotent stem cells derived from mammalian species is not a human ES cell, and the second mammalian species is not a human, the method according to any one of claims 1 to 10.
[12]
[claim12]
12.  A method of re-establishing the stem cells with a chimeric forming ability among the same kind, the following steps:  (I) pluripotent from a species of mammal (pluripotent) stem cells or multipotent of the (multipotent) stem cells, co-cultured with high-quality pluripotent stem cells derived from the same type of mammal;  In co-cultures of (ii) Step (i), to form the cell aggregates, and sorting the cell group containing stem cells derived from the pluripotent stem cells or multipotent stem cells;  (Iii) a group of cells of step (ii) co-cultured with host embryo derived from a mammal of the same type;  (Iv) separating the inner cell mass from the step (iii) co-cultured host embryo; and  (V) by the said inner cell mass cloning the stem cells derived from the pluripotent stem cells or multipotent stem cells, re-establishing the stem cells with a chimeric forming ability in allogeneic; comprising, wherein, the mammal the species derived from pluripotent stem cells or multipotent stem cell, or pluripotent stem cells or multipotent stem cells derived from a species of mammals other than rodents, rodent-derived induced pluripotent stem cells (iPS cell) or multipotent stem cells, the method.
[13]
[claim13]
13.  Further, the re-established stem cells in step (v), comprising in conjunction with high-quality pluripotent stem cells derived from the same type of mammal to obtain a population of cells, repeating steps (iii) ~ (v), claims the method according to 12.
[14]
[claim14]
14.  A method of re-establishing the stem cells with a chimeric forming ability among the same kind, the following steps:  (I) pluripotent from a species of mammal (pluripotent) stem cells or multipotent of the (multipotent) stem cells, combined with high-quality pluripotent stem cells derived from a mammal of the same species get a group of cells;  (Ii) a group of cells of step (i), co-cultured with host embryo derived from the same type of mammal;  (Iii) separating the inner cell mass from the step (ii) co-cultured host embryo; and  (Iv) by cloning the stem cells derived from the pluripotent stem cells or multipotent stem cells from the inner cell mass, and re-establishing the stem cells with a chimeric forming ability among the same kind; Including, where, or pluripotent stem cells or multipotent stem cells derived from the mammal species are pluripotent stem cells or multipotent stem cells derived from mammalian species other than rodents, from rodents is the induced pluripotent stem cells (iPS cells) or multipotent stem cells, the method.
[15]
[claim15]
15.  Comprising repeating the steps (i) ~ (v), this time, the second and subsequent cycles step (i) is re-established stem cells in the immediately preceding cycle of step (iv), from the same kind of mammal high performed by obtaining a cell group combined with quality pluripotent stem cells, the method of claim 14.
[16]
[claim16]
16.  Species of mammals is not a human, the method according to any one of claims 12 to 15.
[17]
[claim17]
17.  A stem cell, it is those were re-established from pluripotent stem cells or multipotent stem cells or rodent-derived iPS cells or multipotent stem cells, derived from a species of mammals other than rodents, and, the following:  Chimeric can be formed;  · Cell aggregates allow the formation; and  - The niche environment of inner cell mass has a high affinity; having 1 or more than the features selected the group consisting of said cell.
[18]
[claim18]
18.  The preceding claims 1 was re-established by the method described in 16, cells of claim 17.
[19]
[claim19]
19.  A method of performing efficacy evaluation or condition analysis using cells comprising:  (I) obtaining a cell: Where the cell is a stem cell with a chimeric forming ability was reestablished by the method according to any one of (A) according to claim 1 to 16, (B) Claim 1 to 16 or somatic stem cells were obtained by differentiating the stem cells with a chimeric forming ability was reestablished by the method described in one of a organ progenitor cells or somatic cells, wherein the somatic stem cells , organ progenitor cells or somatic cells, is obtained by any of the following methods (a) ~ (c):  (A) from the stem cells with chimeric forming ability was reestablished by the method according to any one of claims 1 to 16 to produce a chimeric embryo or chimeric fetuses / fetuses, and from the chimeric embryo or chimeric fetuses / fetuses somatic stem cells, organ precursor cell or a somatic cell, get;  (B) (a) a chimeric embryo or chimeric fetus / embryo-derived somatic stem cells obtained in were differentiated in vitro get the organ precursor cell or somatic cells;  (C) By differentiating stem cells with re-established chimeric forming ability in vitro by the method of any one of claims 1 to 16, obtained somatic stem cells, organ progenitor cells or somatic cells;  Step for efficacy evaluation or condition analysis using cells obtained in (ii) Step (i); Including the said method.
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • JICHI MEDICAL UNIVERSITY
  • Inventor
  • ENDO HITOSHI
  • NAGAO YASUMITSU
  • HANAZONO YUTAKA
  • TOMINAGA KAORU
  • OHMORI TSUKASA
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 DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR 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 SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN 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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