CME W1-Cl.8+

Source: DGRC, stock #151. When ordering, please also send an e-mail to the DGRC (dgrc@indiana.edu) stating that you are ordering these cells for modENCODE; this will ensure that you receive cells from the lot that has been reserved for modENCODE.

Medium

M3 medium + 2% heat-inactivated FCS + 5 μg/ml insulin + 2.5% fly extract.

Note:Cl.8 cells are not very sensitive to the lot of FCS. Nonetheless, we recommend that everyone use the same lot of serum (see below) to maximize consistency within modENCODE.

Thawing cells

As soon as possible after you receive them, thaw an ampoule of frozen cells in a 25 cm2 T-flask, precisely as described in the DGRC protocol [https://dgrc.bio.indiana.edu/include/file/ThawingCells.pdf].

Cl.8 cells recover slowly after thawing. Once they reach approximately 107 cells/ml (enough to cover the surface of the flask), they should be diluted as follows: Using a Pasteur pipet, blow medium at the surface of the flask to dislodge the cells; Cl.8 cells are the most strongly surface-adherent of the 4 modENCODE lines, but they can still be dislodged by this procedure. Transfer the contents of the flask to a 10 cm plate; add 5 ml of fresh medium to the plate and pipet gently up and down to mix. Add 5 ml of fresh medium to the flask; there are generally enough cells left on the surface that they will readily grow up. Sometimes, after a few days the cells in the plate do not seem as healthy as those in the flask; if this happens, go back to the flask and repeat the procedure. It will take several transfers for the cells to reach their normal growth rate; they should not be used for experiments during that time.

General instructions for culture maintenance


Citing the DGRC

When publishing experiments using materials obtained from the DGRC please follow the citation guidelines on the material's stock page, including citing the Drosophila Genomics Resource Center, supported by NIH grant 2P40OD010949, in the acknowledgments. Your cooperation helps us when we need to renew our grant as well as the researchers that donate materials to the DGRC.