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.