Beating behavior of primary neonatal cardiomyocytes and cardiac-differentiated P19.CL6 cells on different extracellular matrix components
JOURNAL OF ARTIFICIAL ORGANS
Authors: Miskon, Azizi; Ehashi, Tomo; Mahara, Atsushi; Uyama, Hiroshi; Yamaoka, Tetsuji
Abstract
Stem cell-based therapy in cardiac tissue engineering is an emerging field that shows great potential for treating heart diseases. However, even preliminary issues, such as the ideal niche for cardiomyocytes, have not been clarified yet. In the present study, the effects of extracellular matrix (ECM) components on the beating duration of neonatal rat cardiomyocytes (RCMs) and on the cardiac differentiation of P19.CL6 carcinoma stem cells were studied. RCMs were cultured on gelatin-, fibronectin-, and collagen type I-coated dishes and on noncoated polystyrene dishes, and their beating rate, beating duration, and cardiac gene expression were evaluated. The beating period and the expression of troponin T type-2 (TNNT2) and troponin C type-1 (TNNC1) of cardiomyocytes cultured on gelatin-coated dishes were longer and higher than for those on dishes with other coatings. For the cardiac differentiation of P19.CL6 cells, troponin T type-2 expression on gelatin- and fibronectin-coated dishes was five times that on collagen type I-coated dishes or polystyrene dishes 11days after induction. These results indicate that a gelatin-coated surface has a high ability not only to maintain the cardiac phenotype but also to enhance cardiac differentiation.
Transcriptomic Profile of Primary Culture of Skeletal Muscle Cells Isolated from Semitendinosus Muscle of Beef and Dairy Bulls
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Authors: Ciecierska, Anna; Motyl, Tomasz; Sadkowski, Tomasz
Abstract
The aim of the study was to identify differences in the transcriptomic profiles of primary muscle cell cultures derived from the semitendinosus muscle of bulls of beef breeds (Limousin (LIM) and Hereford (HER)) and a dairy breed (Holstein-Friesian (HF)) (n= 4 for each breed). Finding a common expression pattern for proliferating cells may point to such an early orientation of the cattle beef phenotype at the transcriptome level of unfused myogenic cells. To check this hypothesis, microarray analyses were performed. The analysis revealed 825 upregulated and 1300 downregulated transcripts similar in both beef breeds (LIM and HER) and significantly different when compared with the dairy breed (HF) used as a reference. Ontological analyses showed that the largest group of genes were involved in muscle organ development. Muscle cells of beef breeds showed higher expression of genes involved in myogenesis (includingerbb-3,myf5,myog,des,igf-1,tgfb2) and those encoding proteins comprising the contractile apparatus (acta1,actc1,myh3,myh11,myl1,myl2,myl4,tpm1,tnnt2,tnnc1). The obtained results confirmed our hypothesis that the expression profile of several groups of genes is common in beef breeds at the level of proliferating satellite cells but differs from that observed in typical dairy breeds.