title: A P19 and P19CL6 cell-based complementary approach to determine paracrine effects in cardiac tissue engineering.
abstract: The negligible self-repair potential of the myocardium has led to cell-based tissue engineering approaches to restore heart function. There is more and more consensus that, in addition to cell development, paracrine effects in particular play a pivotal role in the repair of heart tissue. Here, we present two complementary murine P19 and P19CL6 embryonic carcinoma cell-based in vitro test approaches to study the potential of repair cells and the factors secreted by these cells to induce cardiomyogenesis. P19 cells were 3-dimensionally cultured in hanging drops and P19CL6 cells in a monolayer. Both systems, capable of inducible differentiation towards the cardiomyogenic lineage shown by the appearance of beating cells, the expression of connexin 43 and cardiac troponins T and I, were used to test the cardiomyogenesis-inducing potential of human cardiac-derived adherent proliferating (CardAP) cells, which are candidates for heart repair. CardAP cells in coculture as well as CardAP cell-conditioned medium initiated beating in P19 cells, depending on the cell composition and concentration of the medium. CardAP cell-dependent beating was not observed in P19CL6 cultures, but connexin 43 and cardiac troponin formation as well as expression of GATA-binding protein 4 indicated the dose-dependent stimulatory cardiomyogenic effect of human CardAP cells. In summary, in different ways, P19 and P19CL6 cells have shown their capability to detect paracrine effects of human CardAP cells. In a complementary approach, they could be beneficial for determining the stimulatory cardiomyogenic potential of candidate cardiac-repair cells in vitro.
authors.affiliation: Berlin-Brandenburg Center for Regenerative Therapies, Charité-Universitätsmedizin Berlin, Berlin, Germany.
authors.forename: Tilo
authors.initials: T
authors.lastname: Dehne
authors.forename: Xavier
authors.initials: X
authors.lastname: Adam
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Materne
authors.forename: Mareike Carola
authors.initials: MC
authors.lastname: Reimann
authors.forename: Jan Philipp
authors.initials: JP
authors.lastname: Krüger
authors.forename: Sophie
authors.initials: S
authors.lastname: Van Linthout
authors.forename: Carsten
authors.initials: C
authors.lastname: Tschöpe
authors.forename: Marion
authors.initials: M
authors.lastname: Haag
authors.forename: Michael
authors.initials: M
authors.lastname: Sittinger
authors.forename: Jochen
authors.initials: J
authors.lastname: Ringe
mesh_terms: D000818:Animals; D018236:Carcinoma, Embryonal; D002454:Cell Differentiation / Q000502:physiology; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D006321:Heart / Q000502:physiology*; D006801:Humans; D051379:Mice; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D017077:Culture Media, Conditioned