Method for generating T cell lineage populations from stem/progenitor cells
AssigneeNotch Therapeutics (Canada) Inc.
InventorsRichard Carpenedo, Alessia Pallaoro, Steven Woodside
Life of this patent
Estimating the term…
Abstract
The present disclosure provides a method for generating a T cell lineage population from progenitor T cells. The method comprises culturing the progenitor cells in the presence of Notch signalling ligand, such as Delta-like-4 (DL4), provided on a surface area of at least 7 square centimetres per millilitre culture volume (7 cm2/mL). Population of cells produced using the method are provided as well as methods of using same. Further provided is a method of differentiating a progenitor T cell population enriched for CD4−CD8+ TCRγδ+ cells comprising culturing progenitor T cells in the presence of Notch signalling ligand, such as DL4, provided on a surface area of 0.78 to 4.7 cm2/mL.
Claims24
A method of generating a human T cell lineage cell population, the method comprising:
- a) providing a population of human progenitor T cells;
- and
- b) culturing the progenitor T cells in the presence of a surface-bound recombinant Notch ligand Delta-like-4 (DL4),
- wherein the recombinant DL4 is provided on a culture plate, one or more particles, or one or more beads;
- wherein the recombinant DL4 is provided on a surface area of between 7 square centimetres per millilitre culture volume (7 cm2/mL) and 56 cm2/mL;
- wherein the concentration of the recombinant DL4 is between 7.89×1010 and 1.66×1013 molecules/mL culture volume;
- wherein the culturing is for at least 3 days;
- wherein the progenitor T cells comprise CD5+ CD7+ cells;
- wherein the T cell lineage cell population comprises CD4− CD8+ cells;
- and wherein the CD4− CD8+ cells are generated from the CD5+ CD7+ progenitor T cells, bypassing a CD4+ CD8+ cell stage.
A method of generating a human T cell lineage cell population, the method comprising:
- a) providing a population of human progenitor T cells;
- and
- b) culturing the progenitor T cells in a culture vessel in the presence of a recombinant DL4,
- wherein the recombinant DL4 is provided on a three-dimensional substrate, and wherein the ratio of the surface area of the substrate to a culturing surface area of the culture vessel is between 1.77 to 1 (1.77:1) and 14 to 1 (14:1);
- wherein the concentration of the recombinant DL4 is between 7.89×1010 and 1.66×1013 molecules/mL culture volume;
- wherein the culturing is for at least 3 days;
- wherein the progenitor T cells comprise CD5+ CD7+ cells;
- wherein the T cell lineage cell population comprises CD4− CD8+ cells;
- and wherein the CD4− CD8+ cells are generated from the CD5+ CD7+ progenitor T cells, bypassing a CD4+ CD8+ cell stage.
Dependencies are inferred from the claim text.