公开(公告)号：US20230374458A1

公开(公告)日：2023-11-23

申请号：US18023289

申请日：2021-08-25

Applicant: UNIVERSITY HEALTH NETWORK

Inventor： Gordon Keller ,  Michael Atkins

IPC: C12N5/0789

CPC classification number: C12N5/0647 ,  C12N2506/45 ,  C12N2501/115 ,  C12N2501/155 ,  C12N2501/16 ,  C12N2501/724 ,  C12N2501/42

Abstract: Provided are methods for making yolk sac like hematopoietic progenitors by specifying a KDR+CD235a/b+ mesoderm cells capable of giving rise to T lymphoid lineage cells or cells differentiated therefrom. The method involves contacting pluripotent stem cells (PSCs) with a mesoderm specifying culture composition comprising a BMPR1/R2 agonist, an FGF receptor agonist and an activin receptor agonist to produce a KDR+CD235a/b+ mesoderm cells; and optionally isolating the KDR+CD235a/b+ mesoderm cells.

公开(公告)号：US20220025324A1

公开(公告)日：2022-01-27

申请号：US17277637

申请日：2019-09-18

Applicant: University Health Network

Inventor： Gordon Keller ,  Blair Kenneth Gage

IPC: C12N5/071

Abstract: Disclosed herein are methods of producing a population of venous angioblast cells from stem cells using a venous angioblast inducing media and optionally isolating a CD34+ population from the cell population comprising the venous angioblast cells, for example using a CD34 affinity reagent, CD31 affinity reagent and/or CD144 affinity reagent, optionally with or without a CD73 affinity reagent as well as methods of further differentiating the venous angioblasts in vitro to produce SEC-LCs and/or in vivo to produce SECs. Uses of the cells and compositions comprising the cells are also described.

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.

公开(公告)号：US20240398869A1

公开(公告)日：2024-12-05

申请号：US18526459

申请日：2023-12-01

Applicant: University Health Network

Inventor： Gordon Keller ,  Stephanie Protze ,  Jee Hoon Lee

IPC: A61K35/28 ,  A61K35/34 ,  A61P9/04 ,  C12N5/077 ,  G01N33/50 ,  G01N33/68

Abstract: Methods are disclosed for producing populations of cardiomyocytes from pluripotent stem cells. Populations may be enriched for either atrial or ventricular cardiomyocytes and the resulting ventricular population may be essentially free of pacemaker cells. The method includes incubating pluripotent stem cells in a suitable medium with a BMP component, and an activin component, the amounts of activin may be varied to enrich for either atrial or ventricular cardiomyocytes. The enriched populations, as well as methods of using the same to treat patients in need of cardiac repair are disclosed.

公开(公告)号：US20210062153A1

公开(公告)日：2021-03-04

申请号：US16900970

申请日：2020-06-14

Applicant: University Health Network

Inventor： Gordon Keller ,  Alec Drake Witty ,  Steven James Kattman

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

Abstract: Methods and products for obtaining cardiovascular lineage cells from hPSCs. The method comprises one or more of the following steps: (a) contacting BMP component primed hPSCs with a cardiovascular mesoderm programming cocktail and culturing the contacted hPSCs for a period of time to generate a KDR+ and PDGFRalpha+ cardiovascular mesoderm cell population; (b) contacting the cardiovascular mesoderm cell population with a cardiovascular progenitor specification cocktail and culturing the contacted cardiovascular mesoderm cell population for a period of time to generate a NKX2-5+ or WT1+ cardiovascular progenitor cell population; and (c) contacting the cardiovascular progenitor cell population with a maturation cocktail and culturing the contacted cardiovascular progenitor population for a period of time to produce a cardiovascular population optionally cardiomyocyte lineage cells expressing cardiac troponin T (cTnT) and/or SIRPA and/or epicardial lineage cells expressing WT1.

公开(公告)号：US10711246B2

公开(公告)日：2020-07-14

申请号：US14915992

申请日：2014-09-12

Applicant: UNIVERSITY HEALTH NETWORK

Inventor： Gordon Keller ,  Alec Drake Witty ,  Steven James Kattman

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

Abstract: Provided are methods and products for obtaining cardiovascular lineage cells from hPSCs. The method for obtaining a cardiomyocyte lineage or an epicardial lineage cell population from human pluripotent stem cells (hPSCs) comprises one or more of the following steps: (a) contacting BMP component primed hPSCs with a cardiovascular mesoderm programming cocktail suitable for inducing the hPSCs to differentiate to a cardiovascular mesoderm cell population under conditions suitable for the programming cocktail to penetrate the hPSCs and culturing the contacted hPSCs for a period of time to generate a KDR+ and PDGFRalpha+ cardiovascular mesoderm cell population; (b) contacting the cardiovascular mesoderm cell population with a cardiovascular progenitor specification cocktail suitable to specify a NKX2-5+ or WT1+ cardiovascular progenitor cell population under conditions suitable for the specification cocktail to penetrate the cardiovascular mesoderm cell population and culturing the contacted cardiovascular mesoderm cell population for a period of time to generate a NKX2-5+ or WT1+ cardiovascular progenitor cell population; and (d) contacting the cardiovascular progenitor cell population with a maturation cocktail under conditions suitable for the maturation cocktail to penetrate the cardiovascular progenitor cell population and culturing the contacted cardiovascular progenitor population for a period of time to produce a cardiovascular population optionally cardiomyocyte lineage cells expressing cardiac troponin T (cTnT) and/or SIRPA and/or epicardial lineage cells expressing WT1.

公开(公告)号：US20140322808A1

公开(公告)日：2014-10-30

申请号：US14359517

申请日：2012-11-21

Applicant: University Health Network ,  Sunnybrook Research Institute

Inventor： Gordon Keller ,  Juan Carlos Zuniga-Pflucker ,  Marion J. Kennedy ,  Christopher Michael Sturgeon ,  Andrea Ditadi ,  Geneve Sheandra Awong

IPC: C12N5/0789

CPC classification number: C12N5/0647 ,  C12N2500/24 ,  C12N2500/35 ,  C12N2500/38 ,  C12N2501/105 ,  C12N2501/115 ,  C12N2501/125 ,  C12N2501/14 ,  C12N2501/145 ,  C12N2501/155 ,  C12N2501/16 ,  C12N2501/165 ,  C12N2501/2303 ,  C12N2501/2306 ,  C12N2501/2307 ,  C12N2501/2311 ,  C12N2501/26 ,  C12N2502/1394 ,  C12N2506/02 ,  C12N2506/45 ,  G01N33/56972 ,  G01N2333/70596

Abstract: There is described herein a method of enriching a population of stem cells for hematopoietic progenitors. The method comprises inducing hematopoietic differentiation in a population of human embryonic stem cells or human induced pluripotent stem cells; sorting the population based on expression of CD43 and at least one of CD34, CD31 and CD144; and selecting a fraction that is at least one of CD34+CD43−, CD31+CD43− and CD144+CD43−. Also provided are populations of hematopoietic progenitors obtained by the methods described herein.

Abstract translation: 这里描述了一种富集造血祖细胞群的方法。 该方法包括在人胚胎干细胞或人诱导的多能干细胞群体中诱导造血分化; 基于CD43和CD34，CD31和CD144中的至少一种的表达对群体进行排序; 并选择至少一种CD34 + CD43-，CD31 + CD43-和CD144 + CD43-的分数。 还提供了通过本文所述方法获得的造血祖细胞群。