MAb C19 targets a novel surface marker for the isolation of human cardiac progenitor cells from human heart tissue and differentiated hESCs. (May 2015)

• Record Type: Journal Article
• Authors: Leung, Hau Wan; Moerkamp, Asja T.; Padmanabhan, Jayanthi; Ng, Sze-Wai; Goumans, Marie-José; Choo, Andre
• Is Part Of: Journal of molecular and cellular cardiology. Volume 82(2015:May)
• Page Start: 228
• Page End: 237
• Publication Date: 2015-05
• Subjects: CPC cardiac progenitor cell -- mAb monoclonal antibody -- hESC human embryonic stem cell -- MACS magnetic-activated cell sorting -- EB embryoid body -- Sca-1 stem cell antigen-1 -- KDR kinase domain receptor -- TGFβ1 transforming growth factor beta 1 -- cTnI cardiac troponin I -- cTnT cardiac troponin T; Resident cardiac progenitor cells -- Sca-1 -- hESC differentiation -- Monoclonal antibodies -- Cardiomyocyte differentiation -- Multipotent; Cardiology -- Periodicals; Heart Diseases -- Periodicals; Molecular Biology -- Periodicals; Cardiologie -- Périodiques; Cardiology; Electronic journals; Periodicals; 616.12
• DOI: 10.1016/j.yjmcc.2015.02.016
• Languages: English
• ISSNs: 0022-2828

Abstract

Abstract: Aims: Cardiac progenitor cells (CPCs) have been isolated from adult and developing hearts using an anti-mouse Sca-1 antibody. However, the absence of a human Sca-1 homologue has hampered the clinical application of the CPCs. Therefore, we generated novel monoclonal antibodies (mAbs) specifically raised against surface markers expressed by resident human CPCs. Here, we explored the suitability of one of these mAbs, mAb C19, for the identification, isolation and characterization of CPCs from fetal heart tissue and differentiating cultures of human embryonic stem cells (hESCs). Methods & results: Using whole-cell immunization, mAbs were raised against Sca-1 + CPCs and screened for reactivity to various CPC lines by flow cytometry. mAb C19 was found to be specific for Sca-1 + CPCs, with high cell surface binding capabilities. mAb C19 stained small stem-like cells in cardiac tissue sections. Moreover, during differentiation of hESCs towards cardiomyocytes, a transient population of cells with mAb C19 reactivity was identified and isolated using magnetic-activated cell sorting. Their cell fate was tracked and found to improve cardiomyocyte purity from hESC-derived cultures. mAb C19 + CPCs, from both hESC differentiation and fetal heart tissues, were maintained and expanded in culture, while retaining their CPC-like characteristics and their ability to further differentiate into cardiomyocytes by stimulation with TGFβ1. Finally, gene expression profiling of these mAb C19 + CPCs suggested a highly angiogenic nature, which was further validated by cell-based angiogenesis assays. Conclusion: mAb C19 is a new surface marker for the isolation of multipotent CPCs from both human heart tissues and differentiating hESCs. Highlights: mAb C19 is a novel mAb generated against surface epitopes on resident Sca-1 + CPC. mAb C19 could isolate CPCs from both human heart tissue and differentiated hESCs. mAb C19 + CPCs retained high growth rates and cardiomyocyte differentiation capacity. Gene expression microarrays revealed the angiogenic nature of mAb C19 + CPCs. In vitro differentiation demonstrated multipotency to all three cardiac cell types.