Use of LIFR or FGFR3 as a cell surface marker for isolating human cardiac ventricular progenitor cells
First Claim
1. A method for isolating human cardiac ventricular progenitor cells, the method comprising:
- 1) providing a culture of human embryonic stem (ES) cells or induced pluripotent stem cells (iPSCs);
2) at day 0, activating Wnt/β
-catenin signaling in said culture from step 1;
3) at day 3-5, inhibiting Wnt/β
-catenin signaling in said culture from step 2 to generate human cardiac progenitor cells (CPCs);
4) at day 5-7, contacting said human CPCs from step 3 with one or more agents reactive with Leukemia Inhibitory Factor Receptor (LIFR) and/or Fibroblast Growth Factor Receptor 3 (FGFR3);
5) separating LIFR+ cells and/or FGFR3+ cells from negative cells; and
6) isolating the LIFR+ cells and/or FGFR3+ cells as LIFR+ and/or FGFR3+ human cardiac ventricular progenitor cells.
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Abstract
The present invention provides LIFR and FGFR3 as cell surface markers for isolating human cardiomyogenic ventricular progenitor cells, in particular progenitor cells that preferentially differentiate into cardiac ventricular muscle cells. Thus, the invention provides human ventricular progenitor (HVP) cells. The invention provides in vitro methods of the separation of Islet 1+ LIFR+ ventricular progenitor cells and/or Islet 1+/FGFR3+ ventricular progenitor cells and/or Islet 1+/LIFR+/FGFR3+ ventricular progenitor cells, and the large scale expansion and propagation thereof. Large clonal populations of isolated LIFR+ and/or FGFR3+ ventricular progenitor cells are also provided. Methods of in vivo use of LIFR+ and/or FGFR3+ ventricular progenitor cells for cardiac repair or to improve cardiac function are also provided. Methods of using the LIFR+ and/or FGFR3+ ventricular progenitor cells for cardiac toxicity screening of test compounds are also provided.
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Citations
12 Claims
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1. A method for isolating human cardiac ventricular progenitor cells, the method comprising:
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1) providing a culture of human embryonic stem (ES) cells or induced pluripotent stem cells (iPSCs); 2) at day 0, activating Wnt/β
-catenin signaling in said culture from step 1;3) at day 3-5, inhibiting Wnt/β
-catenin signaling in said culture from step 2 to generate human cardiac progenitor cells (CPCs);4) at day 5-7, contacting said human CPCs from step 3 with one or more agents reactive with Leukemia Inhibitory Factor Receptor (LIFR) and/or Fibroblast Growth Factor Receptor 3 (FGFR3); 5) separating LIFR+ cells and/or FGFR3+ cells from negative cells; and 6) isolating the LIFR+ cells and/or FGFR3+ cells as LIFR+ and/or FGFR3+ human cardiac ventricular progenitor cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification