Cardiac bodies: clusters of spontaneously contracting cells for regenerating cardiac function
First Claim
1. A composition of cardiac bodies, wherein a cardiac body is defined as a cluster of cells that undergoes spontaneous contraction, and contains a majority of cells that express cardiac troponin I (cTnI), cardiac troponin T (cTnT), or atrial natriuretic factor (ANF), or α
- -cardiac myosin heavy chain (MHC) from an endogenous gene; and
wherein the composition is obtainable by a process comprising;
a) differentiating cells from a pPS cell line obtained from a human blastocyst into a cell population in which at least 20% of the cells express cTnI, cTnT, ANF, or MHC from an endogenous gene, b) separating cells that are present in the population as single cells from cells that are present as clusters;
c) resuspending the cells present as clusters in nutrient medium;
d) reculturing the resuspended cells in the nutrient medium; and
e) collecting and washing the recultured cells;
thereby obtaining a composition of cardiac bodies that undergo spontaneous contraction, and contain a majority of cells that express cTnI, cTnT, ANF, or MHC from an endogenous gene.
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Abstract
This disclosure describes clusters of cardiomyocyte lineage cells referred to as cardiac bodies. They can be obtained by differentiating human embryonic stem cells into cells that express cardiomyocyte markers, and separating cells according to their density. Single suspended cells are removed, leaving self-aggregating clusters that can be propagated and enriched in further separation steps. The resulting cardiac bodies express cardiomyocyte markers at levels ˜100-fold above the starting cell population, and undergo spontaneous periodic contraction. The clusters can be used intact or dispersed into single-cell suspensions for use in research, drug screening or the preparation of pharmaceutical compositions for the treatment of cardiac disease.
58 Citations
16 Claims
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1. A composition of cardiac bodies,
wherein a cardiac body is defined as a cluster of cells that undergoes spontaneous contraction, and contains a majority of cells that express cardiac troponin I (cTnI), cardiac troponin T (cTnT), or atrial natriuretic factor (ANF), or α - -cardiac myosin heavy chain (MHC) from an endogenous gene; and
wherein the composition is obtainable by a process comprising;
a) differentiating cells from a pPS cell line obtained from a human blastocyst into a cell population in which at least 20% of the cells express cTnI, cTnT, ANF, or MHC from an endogenous gene, b) separating cells that are present in the population as single cells from cells that are present as clusters;
c) resuspending the cells present as clusters in nutrient medium;
d) reculturing the resuspended cells in the nutrient medium; and
e) collecting and washing the recultured cells;
thereby obtaining a composition of cardiac bodies that undergo spontaneous contraction, and contain a majority of cells that express cTnI, cTnT, ANF, or MHC from an endogenous gene. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
- -cardiac myosin heavy chain (MHC) from an endogenous gene; and
Specification