Patent ID: 8846396
Filing Date: 2014-09-30
Classification: A61K,A61P,C12N

Abstract:
1. A method for isolating cardiospheres from a cardiac tissue biopsy sample, said method comprising the following steps: fragmenting a cardiac tissue biopsy sample to generate a plurality of cardiac tissue fragments, wherein the cardiac tissue biopsy sample is a non-embryonic cardiac tissue obtained from the ventricle, atrium or auricle of a heart; removing non-cardiac tissue and non-cardiac cells from said cardiac tissue fragments by partial enzymatic digestion; culturing said partially digested cardiac tissue fragments in a first culture media until phase-bright cells migrate from said partially digested cardiac tissue fragments; collecting said phase-bright cells; culturing said phase-bright cells in a second culture media on a polylysine treated surface to generate one or more cardiospheres; wherein said cardiospheres are spheroid structure of about 20 μm to about 150 μm in culture, wherein said cardiospheres are multicellular aggregates comprising stem cells, cardiac cells, and endothelial cells, wherein said stem cells express one or more stem cell markers selected from the group consisting of CD-34, ckit, and sca-1 within 12 hours of said cardiospheres being generated, wherein said endothelial cells express one or more endothelial markers selected from the group consisting of KDR, flk-1 and CD31 within 12 hours of said cardiospheres being generated; and harvesting said one or more cardiospheres.