Has anyone successfully cultured cells with flask coating (BSA, collagen and fibronectin)? How can we coat flask with this mixture, what is the coating incubation time? What is the storage conditions of precoated flasks?
Hi Gizem,
I assume that you are working with plastic tissue culture flasks, right?
In that case, you can coat the flasks with a protein solution beween 1-10 ug/ml for 1hr at 25oC.
You have to be careful in keeping the protein solution sterile, by either filtering through a 0.22 micron filter. After the incubation, wash out the flasks with strile PBS containing antibiotics (pen/strep).
You can store the coated flasks with PBS containing pen/strep at +4oC for months.
Just keep an eye out for mold growth
Gizem,
we use fibronectin to coat plastic dishes, usually at the concentration 1-5 μg/cm2. The solution we buy is sterile, ready-to-use. Just pour the solution onto plastic dish and leave it under hood to dry completely. It takes about 30-40 min. The coated culture ware can be stored for 2-4 weeks at 4°C in a closed sterile container. Good luck!
Hi Gizem, I used collagen and fibronectin, in the firsr case I put collagen remove immediately and wait to dry the surface (I have collagen in acetic acid) and store a RT, for the fibronectin I incubate for 2h in the incubator, remove the excess, dry under the hood and store at 20°C. You can find different protocol and are all correct.
The TPP Flasks are really useful in this situation
http://www.sigmaaldrich.com/catalog/product/sigma/z707627?lang=it®ion=IT
http://www.helena-biosciences.com/en/tpp/tissue-culture-flasks-recloseable/
Sometimes I find the flask a little awkward to coat respect the dish!!!
Yes, you can. You can coat to the surfaces by BSA during 1h at RT or overnight at +4C .
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