title: Differential Expression Levels of Integrin α6 Enable the Selective Identification and Isolation of Atrial and Ventricular Cardiomyocytes.
abstract: BACKGROUND
  Central questions such as cardiomyocyte subtype emergence during cardiogenesis or the availability of cardiomyocyte subtypes for cell replacement therapy require selective identification and purification of atrial and ventricular cardiomyocytes. However, current methodologies do not allow for a transgene-free selective isolation of atrial or ventricular cardiomyocytes due to the lack of subtype specific cell surface markers.
  
  
  RESULTS
  In order to develop cell surface marker-based isolation procedures for cardiomyocyte subtypes, we performed an antibody-based screening on embryonic mouse hearts. Our data indicate that atrial and ventricular cardiomyocytes are characterized by differential expression of integrin α6 (ITGA6) throughout development and in the adult heart. We discovered that the expression level of this surface marker correlates with the intracellular subtype-specific expression of MLC-2a and MLC-2v on the single cell level and thereby enables the discrimination of cardiomyocyte subtypes by flow cytometry. Based on the differential expression of ITGA6 in atria and ventricles during cardiogenesis, we developed purification protocols for atrial and ventricular cardiomyocytes from mouse hearts. Atrial and ventricular identities of sorted cells were confirmed by expression profiling and patch clamp analysis.
  
  
  CONCLUSIONS
  Here, we introduce a non-genetic, antibody-based approach to specifically isolate highly pure and viable atrial and ventricular cardiomyocytes from mouse hearts of various developmental stages. This will facilitate in-depth characterization of the individual cellular subsets and support translational research applications.
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Anne Maria
authors.initials: AM
authors.lastname: Wiencierz
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Manuel
authors.initials: M
authors.lastname: Kernbach
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Josephine
authors.initials: J
authors.lastname: Ecklebe
authors.affiliation: Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn, Germany.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Monnerat
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Tomiuk
authors.affiliation: Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Raulf
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Christalla
authors.affiliation: Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Malan
authors.affiliation: Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Hesse
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bosio
authors.affiliation: Institute of Physiology I, Life and Brain Center, University of Bonn, Bonn, Germany.
authors.forename: Bernd K
authors.initials: BK
authors.lastname: Fleischmann
authors.affiliation: Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Eckardt
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018507:Gene Expression Regulation, Developmental; D006325:Heart Atria / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D006352:Heart Ventricles / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D039503:Integrin alpha6 / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D032383:Myocytes, Cardiac / Q000378:metabolism*; D018994:Myosin Light Chains / Q000235:genetics / Q000378:metabolism
chemical_list: D039503:Integrin alpha6; C524495:Mlc2a protein, mouse; D018994:Myosin Light Chains