Hello everyone,
I am trying to research c2c12 differentiation.
It was fine after changing medium( 10% fbs to 2%hs) and c2c12 transformed into myotube in five days.
However, myotube was surrounded by many "small cells" in the end (like the picture in the below).
Is this a other cells in my medium?
*By the way, Is it necessary to remove complement in my HS?
Thanks for your help!
could you explain how do you culture the cells and differntiate them exactly in which kind of culture plate? i can see long myotubes but lots of round shape cells which probably are dead cells
Sahar Salehi-Müller
Thank you for your comment.
I cultured C2C12 cells in 6 well plate with 3 ml medium when I differntiated them (Before the differentiation, I used 75cm² flask with 15 ml medium).
I think it could be the lack of nutrient or not because I seldom saw the small cells in the 75cm² flask.
hello,
its always happen but I don't understand why?
how we can reduce the dead cell in the background?
and what the most proper why to culture these cells (number, sizes of the plate, time of differentiation)
if someone any idea, please I will appreciate the answer.
thanks so much
Nour Al Zaeed
How long after your myotube transformed?
In this web, it says the myotube will degrade after 5~6 days.
https://jikken110.com/protocol31.html
Original text:
Day 5-6でだいたいmyotueの形成がピークに達しますが、使っている試薬などの影響で多少前後するかもしれません。 Myotubeの形成がピークに達した後、徐々にmyotubeの構造が崩壊していきます。
Wei yu Yeh thanks for the answer
I put the cells 2 days in growth medium after that I change it to differentiation medium FBS 2% until 7 days I collected the cells
but when I add refresh medium within those days I feel the dead cells increased? put all people refresh the medium if we don't do that what happens (i think nothing)?