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    METHODS AND COMPOSITIONS FOR GENERATING CHONDROCYTE LINEAGE CELLS AND/OR CARTILAGE LIKE TISSUE
    1.
    发明申请
    METHODS AND COMPOSITIONS FOR GENERATING CHONDROCYTE LINEAGE CELLS AND/OR CARTILAGE LIKE TISSUE 审中-公开
    用于产生肝细胞线粒细胞和/或软骨样组织的方法和组合物

    公开(公告)号：WO2014161075A1

    公开(公告)日：2014-10-09

    申请号：PCT/CA2014/000312

    申请日：2014-04-02

    Applicant: UNIVERSITY HEALTH NETWORK

    Inventor： KELLER, Gordon ,  CRAFT, April M.

    IPC: C12N5/077 ,  A61K35/32 ,  A61P19/00 ,  C12N5/073 ,  C12Q1/02 ,  G01N33/48 ,  G01N33/50

    CPC classification number: A61K35/32 ,  C12N5/0655 ,  C12N2501/115 ,  C12N2501/155 ,  C12N2501/16 ,  C12N2501/415 ,  C12N2501/999 ,  C12N2503/02 ,  C12N2506/02 ,  C12N2506/03 ,  G01N33/5044

    Abstract: A method for generating chondrocytes and/or cartilage, optionally articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue and/or hypertrophic chondrocyte like cells and/or cartilage like tissue, the method comprising: a. culturing a primitive streak-like mesoderm population, optionally a CD56+, PDGFRalpha+ KDR- primitive streak-like mesoderm population, with a paraxial mesoderm specifying cocktail comprising: i. a FGF agonist; ii. a BMP inhibitor; optionally Noggin, LDN-193189, Dorsomorphin; and iii. optionally one or more of a TGFbeta inhibitor, optionally SB431524; and a Wnt inhibitor, optionally DKK1, IWP2, or XAV939; to specify a paraxial mesoderm population expressing cell surface CD73, CD105 and/or PDGFR-beta; b. generating a chondrocyte precursor population comprising: i. culturing the paraxial mesoderm population expressing CD73, CD105 and/or PDGFR-beta at a high cell density optionally in serum free or serum containing media; ii. culturing the high cell density CD73+, CD105+ and/or PDGFRbeta+ paraxial mesoderm population with a TGFbeta3 agonist in serum free media to produce a high cell density Sox9+, collagen 2+ chondrocyte precursor population; and c. either i. culturing the high cell density Sox9+, collagen 2+ chondrocyte precursor population with the TGFbeta3 agonist for an extended period of time to produce an articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue; or ii. culturing the high cell density Sox9+ collagen2+ chondrocyte precursor population with a BMP4 agonist for an extended period of time to produce a hypertrophic chondrocyte like cells and/or cartilage like tissue.

    Abstract translation: 一种用于产生软骨细胞和/或软骨的方法，任选地关节像非增生性软骨细胞和/或软骨样组织和/或肥大软骨细胞样细胞和/或软骨样组织，所述方法包括：a。 培养原始条纹状中胚层群体，任选地具有CD56 +，PDGFRα+ KDR-原始条纹样中胚层群体，其中傍轴中胚层指定混合物，包括：i。 FGF激动剂; II。 BMP抑制剂; 任选地，Noggin，LDN-193189，Dorsomorphin; 和iii。 任选地一种或多种TGFβ抑制剂，任选的SB431524; 和Wnt抑制剂，任选地DKK1，IWP2或XAV939; 指定表达细胞表面CD73，CD105和/或PDGFR-β的近轴中胚层群; 湾 产生软骨细胞前体群体，包括：i。 以高细胞密度任选地在无血清或含血清培养基中培养表达CD73，CD105和/或PDGFR-β的近轴中胚层群; II。 在血清游离培养基中培养具有TGFbeta3激动剂的高细胞密度CD73 +，CD105 +和/或PDGFRβ+近轴中胚层群体以产生高细胞密度Sox9 +，胶原蛋白+2软骨细胞前体种群; 和c。 我是 用TGFbeta3激动剂长时间培养高细胞密度Sox9 +，胶原蛋白+软骨细胞前体种群，以产生关节性非肥大软骨细胞和/或软骨样组织; 或ii。 用BMP4激动剂长时间培养高细胞密度的Sox9 +胶原蛋白2 +软骨细胞前体群体，以产生肥大的软骨细胞样细胞和/或软骨样组织。

    METHODS AND COMPOSITIONS FOR GENERATING CHONDROCYTE LINEAGE CELLS AND/OR CARTILAGE LIKE TISSUE
    5.
    发明公开
    METHODS AND COMPOSITIONS FOR GENERATING CHONDROCYTE LINEAGE CELLS AND/OR CARTILAGE LIKE TISSUE 审中-公开
    产生软骨细胞系和/或软骨组织的方法和组合物

    公开(公告)号：EP2981606A1

    公开(公告)日：2016-02-10

    申请号：EP14778943.2

    申请日：2014-04-02

    Applicant: University Health Network

    Inventor： KELLER, Gordon ,  CRAFT, April M.

    IPC: C12N5/077 ,  A61K35/32 ,  A61P19/00 ,  C12N5/073 ,  C12Q1/02 ,  G01N33/48 ,  G01N33/50

    Abstract: A method for generating chondrocytes and/or cartilage, optionally articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue and/or hypertrophic chondrocyte like cells and/or cartilage like tissue, the method comprising: a. culturing a primitive streak-like mesoderm population, optionally a CD56+, PDGFRalpha+ KDR-primitive streak-like mesoderm population, with a paraxial mesoderm specifying cocktail comprising: i. a FGF agonist; ii. a BMP inhibitor; optionally Noggin, LDN-193189, Dorsomorphin; and iii. optionally one or more of a TGFbeta inhibitor, optionally SB431524; and a Wnt inhibitor, optionally DKK1, IWP2, or XAV939; to specify a paraxial mesoderm population expressing cell surface CD73, CD105 and/or PDGFR-beta; b. generating a chondrocyte precursor population comprising: i. culturing the paraxial mesoderm population expressing CD73, CD105 and/or PDGFR-beta at a high cell density optionally in serum free or serum containing media; ii. culturing the high cell density CD73+, CD105+ and/or PDGFRbeta+ paraxial mesoderm population with a TGFbeta3 agonist in serum free media to produce a high cell density Sox9+, collagen 2+ chondrocyte precursor population; and c. either i. culturing the high cell density Sox9+, collagen 2+ chondrocyte precursor population with the TGFbeta3 agonist for an extended period of time to produce an articular like non-hypertrophic chondrocyte cells and/or cartilage like tissue; or ii. culturing the high cell density Sox9+ collagen2+ chondrocyte precursor population with a BMP4 agonist for an extended period of time to produce a hypertrophic chondrocyte like cells and/or cartilage like tissue.

    Abstract translation: 用于产生软骨细胞和/或软骨，任选关节样非肥厚软骨细胞和/或软骨样组织和/或肥大软骨细胞样细胞和/或软骨样组织的方法，所述方法包括：a。 用近轴中胚层指定混合物培养原条纹样中胚层群体，任选地为CD56 +，PDGFRα+ KDR-原条纹样中胚层群体，其包含：i。 FGF激动剂; II。 BMP抑制剂; 任选Noggin，LDN-193189，Dorsomorphin; 和iii。 任选地一种或多种TGFβ抑制剂，任选SB431524; 和Wnt抑制剂，任选DKK1，IWP2或XAV939; 以指定表达细胞表面CD73，CD105和/或PDGFR-β的旁轴中胚层细胞群; 湾 产生软骨细胞前体群体，所述软骨细胞前体群体包含：i。 在高血球密度下培养表达CD73，CD105和/或PDGFR-β的旁轴中胚层细胞群，任选在无血清培养基或含血清培养基中培养; II。 在无血清培养基中用TGFbeta3激动剂培养高细胞密度CD73 +，CD105 +和/或PDGFRβ+旁轴中胚层群以产生高细胞密度的Sox9 +，胶原2+软骨细胞前体群; 和c。 要么我。 用TGFβ3激动剂培养高细胞密度的Sox9 +，胶原2+软骨细胞前体细胞一段延长的时间以产生关节状的非肥大性软骨细胞和/或类似软骨的组织; 或ii。 用BMP4激动剂培养高细胞密度的Sox9 +胶原2 +软骨细胞前体群延长的时间段以产生肥大的软骨细胞样细胞和/或类似组织的软骨。

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