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MCF7 Breast Cancer Cell Protocol

Product Information

Human MCF7 Breast Cancer Cells

Catalog No. Product Name Product Quantity Storage on Arrival Thawing Instructions Storage once Thawed
Parental MCF7
ax4010 Cells
1 million
Liquid nitrogen Follow protocol N/A
(MCF7/S0.5) cells/vial
Tamoxifen-
ax4011 Resistant MCF7 Cells 1 million cells/vial Liquid nitrogen Follow protocol N/A
(MCF7/TAMR-4)
Tamoxifen-
ax4012 Resistant MCF7 Cells 1 million cells/vial Liquid nitrogen Follow protocol N/A
(MCF7/TAMR-8)
Fulvestrant-
ax4013 Resistant MCF7 Cells 1 million cells/vial Liquid nitrogen Follow protocol N/A
(MCF7/182R-6)
Aliquot & store
ax0044 Unlock 25 mL
at Store at 4°C
-80oC for up to Thaw at 4oC for up to 1
6 months week
Lot-specific information is stated in the Certificate of Analysis for each product.

Recommendations:

  • Recommended culture vessel coating: Not required
  • Recommended cell culture medium: See culture medium section
  • Seeding density (ax4010): 4,000 viable cells/cm2
  • Seeding density (ax4011, ax4012, ax4013): 5,600 viable cells/cm2
  • Recommended centrifugation speed: 200 x g for 5 minutes

Always count the number of viable cells after thawing

Thawing & Plating:

  • Transfer the vial of cells from liquid nitrogen storage with the vial buried in dry ice. Removethe vial from dry ice and transfer it immediately to a 37°C water bath.
  • Thaw the cells quickly in a 37°C water bath until just prior to complete thawing.
  • Wipe the outside of the vial with 70% ethanol.
  • Gently resuspend the cells and transfer to a 15 mL sterile conical tube.
  • Slowly add 10 mL of pre-warmed, 37°C, cell culture medium.
  • Rinse the cryovial with 1 mL of medium to ensure all of the cells are transferred.
  • Centrifuge the cells at 200 x g for 5 min.
  • Carefully remove the supernatant and resuspend the cell pellet in 1-2 mL of pre-warmed, 37°C, cell culture medium.
  • Perform a cell count to determine the number of viable cells.
  • Dilute the cells into the required volume of pre-warmed, 37°C, cell culture medium.
  • Seed cells into the culture vessel at the recommended seeding density.
  • Incubate the cells at 37°C, 5% CO2 in a humidified incubator.
  • Replace the cell culture medium every 2-3 days depending on cell confluency.

Passaging:

  • Passage when the culture reaches: 80-100% confluent
  • Recommended passaging reagent: Unlock
  • Remove all spent medium from cell culture vessels.
  • Gently rinse the surface of the cell layer once with PBS, 2 mL of PBS per 10 cm2 culture surface area. Discard the PBS.
  • Add 1 mL per 10 cm2 of culture surface area of cold/room temperature Unlock passaging reagent. Evenly distribute it over the entire cell layer.
  • Incubate the cells for 5 minutes at 37°C. Observe the cells at regular intervals for detachment from the culture vessel.
  • Once the cells have detached, dilute out the passaging reagent with four volumes pre-warmed, 37°C, cell culture medium. For example, if 1 mL of Unlock is used, then add 4 mL of the medium to stop the reaction.
  • Transfer the cell suspension to a sterile conical tube.
  • Centrifuge the cells at 200 x g for 5 minutes.
  • Carefully remove the supernatant and resuspend the cell pellet in 1-2 mL of pre-warmed, 37°C, cell culture medium.
  • Perform a cell count to determine the number of viable cells.
  • Dilute the cells into the required volume of pre-warmed, 37°C, cell culture medium.
  • Seed cells into the culture vessel at the recommended seeding density.
  • Incubate the cells at 37°C, 5% CO2 in a humidified incubator.

Usage Statement:

Our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans.

The end user is not permitted to transfer or re-freeze the cells. It is recommended that low passage cells are used for endpoint experiments.

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