title: Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
abstract: Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), are capable of differentiating into any cell type in the human body and thus can be used in studies of early human development, as cell models for different diseases and eventually also in regenerative medicine applications. Since the first derivation of hESCs in 1998, a variety of culture conditions have been described for the undifferentiated growth of hPSCs. In this study, we cultured both hESCs and hiPSCs in three different culture conditions: on mouse embryonic fibroblast (MEF) and SNL feeder cell layers together with conventional stem cell culture medium containing knockout serum replacement and basic fibroblast growth factor (bFGF), as well as on a Matrigel matrix in mTeSR1 medium. hPSC lines were subjected to cardiac differentiation in mouse visceral endodermal-like (END-2) co-cultures and the cardiac differentiation efficiency was determined by counting both the beating areas and Troponin T positive cells, as well as studying the expression of OCT-3/4, mesodermal Brachyury T and NKX2.5 and endodermal SOX-17 at various time points during END-2 differentiation by q-RT-PCR analysis. The most efficient cardiac differentiation was observed with hPSCs cultured on MEF or SNL feeder cell layers in stem cell culture medium and the least efficient cardiac differentiation was observed on a Matrigel matrix in mTeSR1 medium. Further, hPSCs cultured on a Matrigel matrix in mTeSR1 medium were found to be more committed to neural lineage than hPSCs cultured on MEF or SNL feeder cell layers. In conclusion, culture conditions have a major impact on the propensity of the hPSCs to differentiate into a cardiac lineage.
authors.affiliation: Institute of Biomedical Technology, University of Tampere, Tampere, Finland.
authors.forename: Marisa
authors.initials: M
authors.lastname: Ojala
authors.forename: Kristiina
authors.initials: K
authors.lastname: Rajala
authors.forename: Mari
authors.initials: M
authors.lastname: Pekkanen-Mattila
authors.forename: Marinka
authors.initials: M
authors.lastname: Miettinen
authors.forename: Heini
authors.initials: H
authors.lastname: Huhtala
authors.forename: Katriina
authors.initials: K
authors.lastname: Aalto-Setälä
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004338:Drug Combinations; D004622:Embryo, Mammalian / Q000166:cytology; D004707:Endoderm / Q000166:cytology; D061252:Feeder Cells; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression / Q000187:drug effects; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D007797:Laminin; D051379:Mice; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D011509:Proteoglycans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055760:SOXF Transcription Factors / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D003470:Culture Media; D004338:Drug Combinations; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; D007797:Laminin; C095552:NKX2-5 protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011509:Proteoglycans; D055760:SOXF Transcription Factors; D014157:Transcription Factors; D016222:Fibroblast Growth Factor 2; C059485:matrigel; D003094:Collagen