These cells are being grown in DMEM with 15% FBS, 1% pen-strep, and 80 uM beta-mercaptoethanol. They take a long time to attach and spread, and grow very slowly. There seems to be an unusual high cell death level (compared to several other lines grown before). Increasing the cell density in seeding seem to improve but they still don't appear very healthy. Any suggestions to improve their health are very welcome.
I have used MIN 6 a long time ago and do not recall any serious issues.
Having said that, not all immortalized cells grow equally vigorously.
Some of my old colleagues used them for quite some time and have published extensively using these cells. If you do a search using MIN-6 cells with Chris Newgard's name, you should have more than enough usable information.
Best...
Usage of β-mercaptoethanol is possibly to reduce differentiation of these cells into something else. On the other hand, β-mercaptoethanol also will affect the structure of most mitogenic cytokines by targeting di-sulphide bonds to result in slower growth.
Try reducing β-mercaptoethanol concentration from 80 to 50uM concentration or can be avoided completely. Include sodium pyruvate if the medium lack it. 10% FBS should be fine. use a splitting ratio of 1:4 - 1:6 depending on the cell growth.
Thank you for all answers. I will certainly check for these references and details.
- Badges
- Science topic