title: Differentiation of human embryonic stem cells to cardiomyocytes on microcarrier cultures.
abstract: We have developed an improved cardiomyocyte differentiation protocol where we stabilized embryoid bodies (EB) in serum- and insulin-free medium (bSFS) supplemented with p38 MAP kinase inhibitor (SB203580) by addition of 10 µm laminin-coated positively charged (protamine sulfate derivatized TSKgel Tresyl-5PW) microcarriers. This protocol achieved a maximum 3-fold cell expansion, differentiation efficiency of 20%, and an overall cardiomyocyte yield of 3 × 10⁵ CM/ml in static conditions. In comparison, EB cultures achieved 1.5-fold cell expansion, differentiation efficiency of 15%, and an overall cardiomyocyte yield of 1.1 × 10⁵ CM/ml. The scalability of this platform was demonstrated in suspended spinner cultures, producing a maximum of 2.14 × 10⁵ CM/ml in 50-ml cultures. This yield is two-fold higher than the control static EB-based platform (1.1 × 10⁵ CM/ml), and seven-fold higher than yields reported in literature, 3.1-9 × 10⁴ CM/ml. The robustness of this protocol was tested with HES-3 and H1 cell lines.
authors.affiliation: Bioprocessing Technology Institute, A*STAR-Agency for Science, Technology and Research, Singapore.
authors.forename: Sherwin
authors.initials: S
authors.lastname: Ting
authors.forename: Marti
authors.initials: M
authors.lastname: Lecina
authors.forename: Shaul
authors.initials: S
authors.lastname: Reuveny
authors.forename: Steve
authors.initials: S
authors.lastname: Oh
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D051379:Mice; D008863:Microspheres; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects
chemical_list: D007797:Laminin