Optimized methods for generation of cardiac stem cells from cardiac tissue and their use in cardiac therapy
AssigneeCapricor, Inc.
InventorsMichelle Kreke, Rachel Smith
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Abstract
The present disclosure relates generally to methods for the increased processing of tissue for the generation of cardiac stem cells, wherein the stem cells are suitable for use in cardiac stem cell therapy. In particular, several embodiments relate to the processing of allogeneic donor cardiac tissue for the generation of multiple patient doses of cardiac stem cells.
Claims20
A method for preparing filtered cardiosphere-derived cells (CDCs) for allogeneic cardiac stem cell therapy, comprising the steps of:
- processing donor cardiac tissue into a plurality of tissue explants ranging from about 100 μm3 to about 800 μm3,
- wherein each of said explants range from about 0.1 to about 0.9 grams;
- enzymatically digesting said explants for a time ranging from about 30 seconds to about 5 minutes;
- culturing said explants at a density of about 1 explant per 400 to 700 cm2 culturing surface area until cells migrate from said explant;
- collecting the cells that migrate from the explant;
- culturing the collected cells to generate cardiospheres (CSps);
- culturing the generated CSps by seeding on a fibronectin-coated dish, plate, or vessel to generate CDCs as a monolayer culture;
- harvesting the generated CDCs;
- and
- filtering said harvested CDCs to remove particles greater than 50 μm in diameter, thereby generating filtered CDCs for allogeneic cardiac stem cell therapy.
A method for preparing filtered cardiosphere-derived cells (CDCs) suitable for allogeneic cardiac stem cell therapy, comprising:
- processing donor cardiac tissue into a plurality of tissue explants, each explant being from about 100 μm3 to about 800 μm3;
- enzymatically digesting said explants;
- culturing, after said digestion, said explants at a density of about 1 explant per 400 to 700 cm2 culturing surface area until cells migrate from said explant;
- collecting the cells that migrate from the explant;
- culturing the collected cells to generate cardiospheres (CSps);
- culturing the generated CSps to generate CDCs as a monolayer culture;
- harvesting the CDCs;
- and
- filtering said harvested CDCs through a first filter to remove particles greater in size than about 140 μm and through a second filter to remove particles greater than about 50 μm, thereby generating filtered CDCs suitable for allogeneic cardiac stem cell therapy.
A method for preparing cardiosphere derived cells (CDCs) for allogeneic cardiac stem cell therapy, comprising:
- dividing donor cardiac tissue according to the region of the heart from which the donor tissue originated;
- processing said donor cardiac tissue into a plurality of cuboidal tissue explants, each explant weighing between about 0.1 to about 0.9 grams;
- culturing said explants until cells migrate from said explant;
- collecting the cells that migrate from the explant;
- culturing the collected cells to generate cardiospheres (CSps);
- culturing the generated CSps by seeding the CSps on a dish, plate, or vessel that generates CDCs as a monolayer culture;
- harvesting the CDCs;
- filtering said harvested CDCs remove particles greater than about 50 μm, thereby generating CDCs for allogeneic cardiac stem cell therapy.
Dependencies are inferred from the claim text.