Use of jagged 1/frizzled 4 as a cell surface marker for isolating human cardiac ventricular progenitor cells

Inventors

Chien, Kenneth R. • Lian, Xiaojun

Assignees

Procella Therapeutics AB

Abstract

The present invention provides Jagged 1 and Frizzled 4 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+ Jagged 1+ ventricular progenitor cells and/or Islet 1+/Frizzled 4+ ventricular progenitor cells and/or Islet 1+/Jagged 1+/Frizzled 4+ ventricular progenitor cells, and the large scale expansion and propagation thereof. Large clonal populations of isolated Jagged 1+ and/or Frizzled 4+ventricular progenitor cells are also provided. Methods of in vivo use of Jagged 1+ and/or Frizzled 4+ ventricular progenitor cells for cardiac repair or to improve cardiac function are also provided. Methods of using the Jagged 1+ and/or Frizzled 4+ ventricular progenitor cells for cardiac toxicity screening of test compounds are also provided.

Core Innovation

The invention relates to a method for isolating human cardiac ventricular progenitor cells from a culture of human embryonic stem cells or induced pluripotent stem cells. The method includes activating Wnt/β-catenin signaling in the culture at day 0 and inhibiting Wnt/β-catenin signaling at day 3-5 to generate human cardiac progenitor cells (CPCs), which are then contacted with one or more agents reactive with Jagged 1.

After contacting the CPCs with Jagged 1 reactivity, Jagged 1+ cells are separated from negative cells and the Jagged 1+ cells are isolated as Jagged 1+ human cardiac ventricular progenitor cells. Dependent variations refine the enrichment by adding an agent reactive with Islet 1 so that Jagged 1+/Islet 1+ cells are separated from negative cells and isolated as Jagged 1+/Islet 1+ human cardiac ventricular progenitor cells.

Further refinement includes obtaining human cardiac ventricular progenitor cells that are Myosin Light Chain 2v (MLC2v) positive. The described approach ties Jagged 1 (JAG1) and, optionally, Islet 1 (ISL1) into the isolation workflow to bias the resulting progenitor population toward ventricular cardiomyocyte differentiation, with downstream differentiation and characterization using ventricular-associated markers such as MLC2v.

The problem addressed is the need to isolate human cardiac ventricular progenitor cells from human embryonic stem cell or induced pluripotent stem cell cultures using marker-based selection. The document frames this as a way to generate an enriched human cardiac ventricular progenitor population suitable for ventricular muscle differentiation and for subsequent use in tissue contexts.

Claims Coverage

The independent claim family member clm-00001 recites 3 major inventive phases: staged Wnt/β-catenin modulation to generate CPCs, marker-reactive Jagged 1-based enrichment, and isolation of Jagged 1+ human cardiac ventricular progenitor cells. Dependent claims clm-00002 through clm-00005 add further inventive refinements by including Islet 1 reactivity, specifying FACS or MACS separation, and adding MLC2v positivity as a qualification for the isolated progenitor cells.

Across the independent claim and its dependents, the claims cover (i) generating CPCs via activating then inhibiting Wnt/β-catenin signaling, and (ii) enriching a ventricular progenitor population using Jagged 1 reactivity, optionally strengthened by Islet 1 dual-marker selection, with further qualification by MLC2v positivity.

Stated Advantages

Documented Applications