TBX5 overexpression stimulates differentiation of chamber myocardium in P19C16 embryonic carcinoma cells.
In vitro differentiation of pluripotent embryonic cells is becoming a model system to study factors and genes involved in early developmental processes including cardiogenesis. An additional application involves the development of donor cells for treatment of diseases among which cardiac infarction. For this purpose differentiated cells should meet the functional characteristics of chamber myocardium, a requirement not convincingly reached as yet. The T-box transcription factor Tbx5 has been demonstrated to be crucial for heart formation. Using stably transfected clones of the P19C16 embryonic carcinoma cell line, reported to differentiate efficiently into the cardiac lineage, we investigated whether Tbx5 is sufficient to enhance cardiogenesis and differentiation of chamber myocardium. TBX5-transfected clones started to beat earlier, however, a relation between transgenic TBX5 mRNA levels and the number of beating foci or levels of Serca2a mRNA, a myocardial marker, could not be observed. However, TBX5-transfected clones displayed significantly higher levels of atrial natriuretic factor (Anf) and Connexin (Cx)40 mRNAs, which are associated with the formation of chamber myocardium. This indicates that Tbx5 enhances cardiac maturation within this system rather than cardiogenesis.
Fijnvandraat
Arnoud C
AC
Experimental and Molecular Cardiology Group, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Lekanne Deprez
Ronald H
RH
Christoffels
Vincent M
VM
Ruijter
Jan M
JM
Moorman
Antoon F M
AF
D000818:Animals; D009320:Atrial Natriuretic Factor / Q000032:analysis / Q000235:genetics; D000252:Calcium-Transporting ATPases / Q000032:analysis / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D017630:Connexins / Q000032:analysis / Q000235:genetics; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D032342:Muscle Cells / Q000737:chemistry / Q000378:metabolism; D009200:Myocardial Contraction / Q000502:physiology; D009206:Myocardium / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D020825:T-Box Domain Proteins / Q000235:genetics* / Q000502:physiology; D014162:Transfection; D000097003:Gap Junction alpha-5 Protein
D009320:Atrial Natriuretic Factor; D000252:Calcium-Transporting ATPases; D017630:Connexins; D004121:Dimethyl Sulfoxide; D012333:RNA, Messenger; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D020825:T-Box Domain Proteins; D000097003:Gap Junction alpha-5 Protein; C103172:T-box transcription factor 5