CELLULES MCF10A HER2 (-/ -),

Application

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.

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°

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

Media renewal changes two to three times per week.

Biochem/physiol Actions

Human epidermal growth factor receptor 2 (HER2) signaling pathway stimulates cell proliferation and survival in the majority of breast cancers. Thus, overexpression of this protein causes breast cancer. Heterodimeric complex of HER2 and phosphatidylinositide 3-kinase (PI3K) is the most potent stimulator of the PI3K/AKT anti-apoptosis pathway. Increased expression of HER2 is also observed in ovarian, colorectal, pancreatic, endometrial and gastric cancers. Trastuzumab, an antibody, works by binding to a domain in the external domain of HER2. This domain is missing in p95, a truncated form of HER2, and hence these cancer cells show resistance to trastuzumab. HER2 protein can be used as a prognostic marker and as a therapeutic option for gynecologic cancers.

Features and Benefits

Zinc finger nuclease (ZFN) knockout on chromosome 17q21.1

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

General description

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

MCF10A CELLS HER2 (-/-) are pre-neoplastic mammary epithelial cells from a human adult female with a ZFN knock out modification.

Quality

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.