Cellular Dynamics International, Inc.

University Health Network

Fujifilm Cellular Dynamics, Inc.

Steven James Kattman

Wen Bo Wang

Casey Stankewicz

Peter Fuhrken

Nathaniel Beardsley

Matt Riley

Gordon Keller

Nathan Meyer

Brad Swanson

Matthew George

Alec Drake Witty

Chad Koonce

Natsuyo Aoyama

Jennifer Luebke-Wheeler

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.

Methods and compositions for generating epicardium cells

Methods and composition for the production of cardiomyocytes from differentiation of pluripotent stem cells are provided. For example, in certain aspects methods including differentiating pluripotent stem cells in a large volume of suspension culture in the presence of ROCK inhibitors are described. In further aspects, methods for differentiation of stem cells into cardiomyocytes that overcome variability between different stem cell clones and different batch of culture medium are provided.

Cardiomyocyte production

Methods and composition for differentiation of pluripotent stem cells are provided. For example, in certain aspects methods including screening of optimal differentiation conditions for a selected stem cell clone in a selected batch of culture medium and use of the determined optimal condition for differentiation into a specific cell lineage.

Titration of differentiation medium components

The present invention concerns methods and compositions for promoting a conversion in stem cells or cells derived therefrom from glycolysis to aerobic respiration and its associated pathways, including oxidative phosphorylation, the TCA cycle, fatty acid oxidation, and pyruvate decarboxylation, for example. In specific embodiments, certain media is employed to improve metabolic maturation of particular stem cell-derived cells, including human embryonic stem cells and human induced pluripotent stem cells, for example. Particular media comprises media without glucose or optionally without pyruvate, with fatty acids, with L-carnitine, with taurine, creatine, non-essential amino acids, L-glutamine, and with anti-oxidants or free-radical scavengers, for example.

Metabolic maturation in stem cell-derived tissue cells

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Madison, WI, United States of America

Steven James Kattman