MDA-MB-453

Frequently Asked Questions

• What are MDA-MB-453 cells, and why should I use them in my research?

  MDA-MB-453 cells are a human breast cancer cell line that originates from a pleural effusion of a patient with metastatic breast cancer. These cells are often used in cancer research.

• What specific growth factors or supplements can enhance the activation of MDA-MB-453 cells?

  Enhancing the activation and growth of MDA-MB-453 cells can be achieved by using specific growth factors and supplements in the culture medium. Here are some key factors and supplements that can be used:

  1. Growth Factors
  Epidermal Growth Factor (EGF): EGF is known to stimulate cell growth and proliferation. Adding EGF to the culture medium can enhance the growth of MDA-MB-453 cells. Typical concentration: 10-20 ng/mL.

  Insulin-like Growth Factor 1 (IGF-1): IGF-1 plays a significant role in cell growth and survival. It can be used to promote the proliferation of breast cancer cells, including MDA-MB-453. Typical concentration: 10-50 ng/mL.

  Heregulin (HRG): Heregulin is a ligand for the HER3 receptor, which can activate HER2 and HER3 signaling pathways. It can be used to enhance growth and signaling in HER2-positive cells. Typical concentration: 10-50 ng/mL.

  2. Hormones
  Androgens (e.g., Dihydrotestosterone, DHT): Since MDA-MB-453 cells express the androgen receptor (AR), adding androgens like dihydrotestosterone (DHT) can activate AR signaling and influence cell growth. Typical concentration: 1-10 nM.

  3. Supplements
  Insulin: Insulin is often added to cell culture media to promote growth and enhance glucose uptake. Typical concentration: 10 µg/mL.

  Hydrocortisone: Hydrocortisone can be used to enhance the growth of certain breast cancer cell lines, including those expressing steroid hormone receptors. Typical concentration: 1 µg/mL.

  Fetal Bovine Serum (FBS): FBS is a common supplement in cell culture media that provides essential growth factors, hormones, and nutrients.Typical concentration: 10-20%.

• How do I passage MDA-MB-453 cells?

  Procedure

  Preparation: Warm the RPMI-1640 medium, PBS, and trypsin-EDTA solution to 37°C in a water bath. Ensure all necessary equipment and reagents are sterile and ready for use.

  Observe Cells: Place the flask containing MDA-MB-453 cells under an inverted microscope.
  Check for cell confluence (typically, cells should be 70-80% confluent before passaging).

  Remove Old Medium: In a biosafety cabinet, aspirate the old medium from the flask carefully without disturbing the cell monolayer.

  Rinse Cells: Add 5-10 mL of PBS to the flask to rinse off any residual medium and serum that may inhibit trypsin activity. Gently rock the flask to ensure the cells are thoroughly rinsed, then aspirate the PBS.

  Add Trypsin-EDTA: Add enough trypsin-EDTA solution to cover the cell monolayer (typically 1-2 mL for a T-75 flask). Gently tilt the flask to ensure the entire surface is covered with trypsin-EDTA.

  Incubate: Place the flask in the incubator for 3-5 minutes, checking the cells under the microscope periodically. The cells should round up and detach from the surface.

  Neutralize Trypsin: Once the cells are detached, add 5-10mL of complete RPMI-1640 medium (with FBS) to the flask to neutralize the trypsin.

  Collect Cells: Gently pipette the cell suspension up and down to ensure all cells are detached and in suspension. Transfer the cell suspension to a 15mL conical tube.

  Centrifuge: Centrifuge the cell suspension at 200×g for 5 minutes to pellet the cells.

  Resuspend Cells: Carefully aspirate the supernatant without disturbing the cell pellet. Resuspend the cells in an appropriate volume of fresh complete RPMI-1640 medium.

  Count Cells (Optional): If needed, take a small aliquot of the cell suspension to count cells using a hemocytometer or an automated cell counter.

  Seed New Flasks: Seed the cells into new culture flasks or plates at the desired density. For example, if splitting at a 1:5 ratio, add 1 part of the cell suspension to 4 parts of fresh medium in a new flask.

  Incubate: Place the newly seeded flasks or plates back into the incubator set at 37°C with 5% CO2.

  Monitoring: Observe the cells under the microscope after 24 hours to ensure they have adhered and are spreading out. Change the medium every 2-3 days or as needed.