MDA-MB-157 Cell Line Derived Xenograft
The utilization of human cells in xenograft tumor models, like the MDA-MB-157 cell lines derived xenograft model (CDX), is imperative in therapeutic development. Breast cancer xenograft models enable investigation of human breast cancer cell therapeutics targeting migration, metastasis, apoptosis, anti-angiogenesis and anti-tumor growth in a human-similar tumor environment. Therapeutic studies in a cell culture setting cannot mimic tumor stroma, vasculature and cell-to-cell interactions as seen in xenograft models.
|MDA-MB-157||ER–, PR–, HER2–, p53(mut)|
|Metastatic Models (Breast)||4T1|
|Non-Metastatic Models (Breast)||BT-474, HS578T, KPL-4, MCF-7, MDA-MB-157, MDA-MB-231, MDA-MB-453, MDA-MB-468, T-47D|
The preparation of tumor xenograft models requires cells growth of large number of tumor cells for inoculation. One million cells usually required for a single injection. Cell population is expanded and obtained by trypsinization from 225 cm2 tissue culture flasks. The collected cells are centrifuged and separated from the supernatant, followed by cell count (via flow cytometry) prior to matrigel injection.
MDA-MB-157 Basic Tumor Xenograft Study Design
- NOD/SCID or Nude (Nu/Nu) mice (8-12 weeks old) are acclimated to the facility for 5-7 days prior to tumor cell injection/implantation.
- Cells to be used for injection are maintained under conditions of exponential growth prior to injection. Cells are prepared for injection by trypsinization and viable cell counts are determined using trypan blue exclusion. Cell count performed by flow cytometry and suspension adjusted to appropriate density. Only cell suspensions with greater than 98% viability are used.
- Each mouse receive one subcutaneous injection in the flank of the hind leg.
- Each site will receive one million tumorigenic cells in a volume of 100 uL containing 50% matrigel (use of matrigel matrix improves tumor cell survival and tumor formation).
- The injection sites are palpated three times weekly until tumors are established. Tumors are measured using digital calipers until they reach an average size of 150-250 mm3. Injected animals are monitored daily with cage-side observations, and body weight measurements.
- Animals are randomized into treatment cohorts and treated according to the treatment schedule. We recommend including cyclophosphamide or other positive control treatment group.
- Once palpable tumors are observed, tumors are measured and mouse weights recorded 2 times a week.
- Animals are euthanized when tumor size reaches 2,000 mm3 size limit. Gross necropsy and tissue collection performed as defined for termination of experiment. Animals showing signs of morbidity due to tumor burden or treatment toxicity are euthanized and processed as defined for termination of experiment.
- Tumors are excised, weighed and documented by digital imaging. Standard necropsies are performed and tissues collected for downstream processing and analysis.