title: Generation of Cardiomyocytes in Pipe-Based Microbioreactor Under Segmented Flow.
abstract: OBJECTIVE
  Embryonic stem (ES) cells have got a broad range differentiation potential. The differentiation is initiated via aggregation of non-differentiated ES cells into embryoid body (EB) capable of multi-lineage development. However experimental variables present in standard differentiation techniques lead to high EB heterogeneity, affecting development into the cells of desired lineage, and do not support the process automatization and scalability.
  
  
  METHODS
  Here we present a novel pipe based microbioreactor (PBM) setup based on segmented flow, designed for spatial maintenance of temperature, nutrition supply, gas supply and sterility.
  
  
  RESULTS
  We verified PBM feasibility for continuous process generating cardiac cells starting from single ES cell suspension followed by EB formation for up to 10 days. The ES cells used in the study were genetically modified for cardiac-specific EGFP expression allowing optical monitoring of cardiomyocytes while EBs remained within PBM for up to 10 days. Efficiency of cardiac cells formation within PBM was similar compared to a standard hanging drop based protocol.
  
  
  CONCLUSIONS
  Our findings ensure further development of microfluidic bioreactor technology to enable robust cardiomyocytes production for needs of drug screening, tissue engineering and other applications.
authors.forename: Dimitry
authors.initials: D
authors.lastname: Spitkovsky
authors.forename: Karen
authors.initials: K
authors.lastname: Lemke
authors.forename: Tobias
authors.initials: T
authors.lastname: Förster
authors.forename: Robert
authors.initials: R
authors.lastname: Römer
authors.forename: Stefan
authors.initials: S
authors.lastname: Wiedemeier
authors.forename: Jürgen
authors.initials: J
authors.lastname: Hescheler
authors.forename: Agapios
authors.initials: A
authors.lastname: Sachinidis
authors.forename: Gunter
authors.initials: G
authors.lastname: Gastrock
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D005434:Flow Cytometry; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D051379:Mice; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods; D008853:Microscopy; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D000071317:Nanog Homeobox Protein / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D023822:Tissue Engineering
chemical_list: D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D012333:RNA, Messenger