Anyone working with mouse fibroblast primary culture? Need to know the culture conditions for the same. Thank you
It's quite straight-forward. 37C in humidified 5% CO2 incubator with 10% FBS media in standard tissue culture plates.
Fibroblast cells can be culture in DMEM medium supplemented with 10% FBS. 37 degree temperature 5% co2 with humidified atmosphere
Thank you Ron Hrstka and Md. Zafaryb. Can you tell me about the passage number? For how many passage do the culture generally survives?
It will depend on what you want to use them for. Also, realize that passage number is not the same as number of population doublings. I have seen people do up to 20 passages but they certainly slow down a lot near the end.
Most of my fibroblast cultures have been for the purpose of making feeders for ES and iPS cells. For that purpose, they become of significantly lower quality after passage 3, which is the passage number I routinely used for feeders. I compared them with P4-6 and there was a progressive reduction in their effectiveness as feeders.
As as absorved, cells divided properly at passage 3. After that cells is not divide properly. As I am working on osteoblast differentiation, I need only 3 passage cells to differentiated into osteoblast cells
For my work, i want to induce fibroblast to myofibroblast and then evaluate with cellular signaling, so the passage number is really important to conduct the many experiments i have to do.
In that case you want to create a large frozen stock of the lowest possible passage number so that you can do many replicates with identical starting material.
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