title: Slow turning lateral vessel bioreactor improves embryoid body formation and cardiogenic differentiation of mouse embryonic stem cells.
abstract: Embryonic stem cells (ESCs) have the ability to form aggregates, which are called embryoid bodies (EBs). EBs mimic early embryonic development and are commonly produced for cardiomyogenesis. Here, we describe a method of EB formation in hydrodynamic conditions using a slow-turning lateral vessel (STLV) bioreactor and the subsequent differentiation of EBs into cardiomyocytes. EBs formed in the STLV were compared with conventional techniques, such as hanging drop (HD) or static suspension cell culture (SSC), for homogeneity of EB size, shape, proliferation, apoptosis, and in vitro cardiac differentiation. After 3 days of culture, a four-fold improvement in the yield of EB formation/mL, a six-fold enhancement in total yield of EB/mL, and a nearly 10-fold reduction of cells that failed to incorporate into EBs were achieved in STLV versus SSC. During cardiac differentiation, a 1.5- to 4.2-fold increase in the area of cardiac troponin T (cTnT) per single EB in STLV versus SSC and HD was achieved. These results demonstrate that the STLV method improves the quality and quantity of ES cells to form EBs and enhances the efficiency of cardiac differentiation. We have demonstrated that the mechanical method of cell differentiation creates different microenvironments for the cells and thus influences their lineage commitments, even when genetic origin and the culture medium are the same. Ascorbic acid (ASC) improved further cardiac commitment in differentiation assays. Hence, this culture system is suitable for the production of large numbers of cells for clinical cell replacement therapies and industrial drug testing applications.
authors.affiliation: 1 BioTalentum Ltd ., Gödöllö, H-2100, Hungary .
authors.forename: Sasitorn
authors.initials: S
authors.lastname: Rungarunlert
authors.forename: Nuttha
authors.initials: N
authors.lastname: Klincumhom
authors.forename: Theerawat
authors.initials: T
authors.lastname: Tharasanit
authors.forename: Mongkol
authors.initials: M
authors.lastname: Techakumphu
authors.forename: Melinda K
authors.initials: MK
authors.lastname: Pirity
authors.forename: Andras
authors.initials: A
authors.lastname: Dinnyes
mesh_terms: D000818:Animals; D001483:Base Sequence; D019149:Bioreactors; D002454:Cell Differentiation; D002478:Cells, Cultured; D017931:DNA Primers; D058732:Embryoid Bodies / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000196:embryology*; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers
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