公开(公告)号：US10736923B2

公开(公告)日：2020-08-11

申请号：US15977536

申请日：2018-05-11

Applicant: UNIVERSITY HEALTH NETWORK

Inventor： Gordon Keller ,  April M. Craft

IPC: C12N5/077 ,  G01N33/50 ,  A61K35/32

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.

公开(公告)号：US20190322986A1

公开(公告)日：2019-10-24

申请号：US15977505

申请日：2018-05-11

Applicant: UNIVERSITY HEALTH NETWORK

Inventor： Gordon Keller ,  April M. Craft ,  Nicole C. Dubois

IPC: C12N5/077 ,  C12Q1/6881

Abstract: The present invention relates to in vitro methods of enriching populations of human pluripotent stem cells that are induced to differentiate to cardiomyocyte progenitor cells and cardiomyocyte cells. The cell populations can be enriched by isolating cells that express SIRPA. The invention also related to in vitro-enriched populations of cardiomyocyte cells and cardiomyocyte progenitor cells obtained from populations of pluripotent stem.

公开(公告)号：US20200239848A1

公开(公告)日：2020-07-30

申请号：US16847597

申请日：2020-04-13

Applicant: University Health Network

Inventor： Gordon Keller ,  April M. Craft ,  Nicole C. Dubois

IPC: C12N5/077 ,  C12Q1/6881 ,  G01N33/569 ,  G01N33/50 ,  G01N33/53

Abstract: The present invention relates to in vitro methods of enriching populations of human pluripotent stem cells that are induced to differentiate to cardiomyocyte progenitor cells and cardiomyocyte cells. The cell populations can be enriched by isolating cells that express SIRPA. The invention also related to in vitro-enriched populations of cardiomyocyte cells and cardiomyocyte progenitor cells obtained from populations of pluripotent stem.

公开(公告)号：US09993504B2

公开(公告)日：2018-06-12

申请号：US14782070

申请日：2014-04-02

Applicant: UNIVERSITY HEALTH NETWORK

Inventor： Gordon Keller ,  April M. Craft

IPC: C12N5/077 ,  A61K35/32 ,  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.