How do you differentiate C2C12 cells?
Differentiation of C2C12 cells is achieved by replacing GM to differentiation media, DM [DMEM—high glucose no sodium pyruvate (Gibco), 2% horse serum (Gibco), 1% glutamine (Gibco), 1% pen/strep (Gibco)]. After 24 h in DM, fused cells should be visible. DM should be changed every 48 h.
Where do C2C12 cells come from?
C2C12 is a subclone from a myoblast line established from normal adult C3H mouse leg muscle1. The original cell line, C2, was obtained by Yaffe and Saxel in 1977 by establishing primary cultures from the thigh muscle of 2 month old normal mice, 70 hours after crush injury2.
How big are C2C12 cells?
At the time of the biophysical studies, mean myotube diameter was 12 microns (range 5-25 microns), and mean length was 290 microns (range 130-520 microns).
Are C2C12 cells adherent?
From the C2s the immortal subline C2C12 was selected (Blau et al., 1985). These cells differentiate well into myocytes under appropriate culture conditions given below. The cells are adherent in culture and are grown on Nunc delta surface plastic culture dishes.
What are L6 myotubes?
The L6 myogenic line was isolated originally by Yaffe from primary cultures of rat thigh muscle maintained for the first two passages in the presence of methyl cholanthrene. Genes expressed myosin Comments. L6 cells fuse in culture to form multinucleated myotubes and striated fibers.
What are Myosatellite cells?
Myosatellite cells, also known as satellite cells, muscle stem cells or MuSCs, are small multipotent cells with very little cytoplasm found in mature muscle. Satellite cells are precursors to skeletal muscle cells, able to give rise to satellite cells or differentiated skeletal muscle cells.
Where can you find Myosatellite cells?
Myosatellite cells are located between the basement membrane and the sarcolemma of muscle fibers, and can lie in grooves either parallel or transversely to the longitudinal axis of the fibre.
Are satellite cells unipotent?
For instance, hematopoietic stem cells are multipotent in that they give rise to all of the differentiated cell types in blood. Satellite cells, by contrast, are unipotent, since they normally give rise to only one type of differentiated progeny, that is, myogenic precursor cells.
Do satellite cells turn into myoblasts?
More specifically, upon activation, satellite cells can re-enter the cell cycle to proliferate and differentiate into myoblasts.
Why do we need satellite cells?
Satellite cells are the source of the new nuclear material that is required for muscle growth and hypertrophy, and if the muscle fiber is damaged, satellite cells become activated, divide, and fuse to replace the damaged portions.
What is the main function of satellite cells?
Satellite cells are precursors to skeletal muscle cells and are responsible for the ability of muscle tissue to regenerate. ie These embryonic cells remain in the adult and can replace damaged muscle fibers to some degree.
How does MDFI-OE promote C2C12 cell differentiation?
We also established the regulatory networks of Mdfi-OE on C2C12 cell differentiation and muscle fiber type transformation and identified hub genes. Further, both RNA-seq and experimental verification demonstrated that Mdfi promoted C2C12 cell differentiation by upregulating the expression of Myod, Myog, and Myosin.
What is the source of C2C12 cells used in the study?
C2C12 cells were purchased from the American Type Culture Collection (ATCC, CRL-1772). All cells used in experiments were passaged less than 10 times from the original plug. C2C12 were authenticated by testing for differentiation efficiency upon receipt. They were not tested for mycoplasma throughout the course of the study.
How are mRNA profiles generated from C2C12 cells?
The mRNA profiles were generated by sequencing six C2C12 cell samples (three each from Mdfi-OE and WT). Sample’s length, clean reads, Q30 quality scores, GC content, and unique mapped rate were summarized in Table 1.
How many cells pass QC in Illumina RNA-Seq data?
The resultant reads were then filtered on which combinations of barcodes were also seen in the Illumina single-cell/nucleus RNA-seq data, which yielded 567 of the 568 cells that passed QC in the Illumina data (Additional file 1: Fig. S2B). The reads were then trimmed of their barcodes to facilitate mapping, and cell identity barcodes were recorded.