MCF10A CELLS ESR2 (-/-)

MCF10A CELLS ESR2 (-/-) are pre-neoplastic mammary epithelial cells from a human adult female with a ZFN knock out modification.

This cell line corresponds to ATCC Catalog No. CRL-10317.

Cell freezing medium-DMSO 1X, Cat. No. C6164

Media renewal changes two to three times per week.

Rapidly thaw vial by gentle agitation in 37°C water bath (~2 minutes), keeping vial cap out of the water. Decontaminate with 70% ethanol, add 9 mL culture media and centrifuge 125 x g (5-7 minutes). Resuspend in complete culture media and incubate at 37°

The base medium for this cell line is Dulbecco′s Modified Eagle′s Medium/Ham′s Nutrient Mixture F-12, Cat. No. 51448C. To make the complete growth medium, add the following components to the base medium: horse serum, Cat No. H1270, to a final concentration (v/v) of 5%; cholera toxin, Cat No. C8052, to a final concentration of 1 ng/mL; human insulin, Cat No. I9278, to a final concentration of 10 ug/mL; epidermal growth factor, Cat No. E9644, to a final concentration of 10 ng/mL; and hydrocortisone, Cat No. H6909, to a final concentration of 0.5 ug/mL.

These MCF10A cells are adherent with a doubling time of approx. 20 hours

Zinc finger nuclease (ZFN) knockout on chromosome 14q23.2

Tested for Mycoplasma, sterility, post-freeze viability, short terminal repeat (STR) analysis for cell line identification, cytochrome oxidase I (COI) analysis for cell line species confirmation.