title: Isolation and expansion of functionally-competent cardiac progenitor cells directly from heart biopsies.
abstract: The adult heart contains reservoirs of progenitor cells that express embryonic and stem cell-related antigens. While these antigenically-purified cells are promising candidates for autologous cell therapy, clinical application is hampered by their limited abundance and tedious isolation methods. Methods that involve an intermediate cardiosphere-forming step have proven successful and are being tested clinically, but it is unclear whether the cardiosphere step is necessary. Accordingly, we investigated the molecular profile and functional benefit of cells that spontaneously emigrate from cardiac tissue in primary culture. Adult Wistar-Kyoto rat hearts were minced, digested and cultured as separate anatomical regions. Loosely-adherent cells that surround the plated tissue were harvested weekly for a total of five harvests. Genetic lineage tracing demonstrated that a small proportion of the direct outgrowth from cardiac samples originates from myocardial cells. This outgrowth contains sub-populations of cells expressing embryonic (SSEA-1) and stem cell-related antigens (c-Kit, abcg2) that varied with time in culture but not with the cardiac chamber of origin. This direct outgrowth, and its expanded progeny, underwent marked in vitro angiogenic/cardiogenic differentiation and cytokine secretion (IGF-1, VGEF). In vivo effects included long-term functional benefits as gauged by MRI following cell injection in a rat model of myocardial infarction. Outgrowth cells afforded equivalent functional benefits to cardiosphere-derived cells, which require more processing steps to manufacture. These results provide the basis for a simplified and efficient process to generate autologous cardiac progenitor cells (and mesenchymal supporting cells) to augment clinically-relevant approaches for myocardial repair.
authors.affiliation: Heart Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA.
authors.forename: Darryl R
authors.initials: DR
authors.lastname: Davis
authors.forename: Eddy
authors.initials: E
authors.lastname: Kizana
authors.forename: John
authors.initials: J
authors.lastname: Terrovitis
authors.forename: Andreas S
authors.initials: AS
authors.lastname: Barth
authors.forename: Yiqiang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Rachel Ruckdeschel
authors.initials: RR
authors.lastname: Smith
authors.forename: Junichiro
authors.initials: J
authors.lastname: Miake
authors.forename: Eduardo
authors.initials: E
authors.lastname: Marbán
mesh_terms: D043925:Angiogenesis Inducing Agents / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001706:Biopsy; D002316:Cardiotonic Agents / Q000378:metabolism; D002454:Cell Differentiation; D019070:Cell Lineage; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D016207:Cytokines / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D009203:Myocardial Infarction / Q000473:pathology / Q000503:physiopathology; D009206:Myocardium / Q000473:pathology*; D032383:Myocytes, Cardiac / Q000166:cytology; D018919:Neovascularization, Physiologic; D010641:Phenotype; D051381:Rats; D017208:Rats, Wistar; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D016276:Ventricular Function / Q000502:physiology
chemical_list: D043925:Angiogenesis Inducing Agents; D015415:Biomarkers; D002316:Cardiotonic Agents; D016207:Cytokines