Mia paca 2 human pancreatic cancer cells

A549 human lung adenocarcinoma epithelial cells, MDA-MB-231 human breast cancer cells, MIA PaCa-2 human pancreatic cancer cells, RKO human colon cancer cells, PC3 human prostate cancer cells, and U87-MG glioblastoma cells were obtained from American Type Culture Collection (ATCC) and cultured in DMEM or RPMI medium. Cells were incubated at 37^oC in a 5% CO₂-containing humidified incubator unless otherwise specified. All cell lines were tested for the presence of mycoplasma via PCR.

Thirty 6-week old male athymic nude mice were purchased from Jackson Laboratories, (Bar Harbor, ME) and placed on chow diet for 1 week following arrival at the UT Austin Animal Resource Center. Mice (n = 15/group) were randomized to receive either the control diet or 30% CR diet used in the previous study. Animals were singly housed and received diet for 23 weeks. Mean body weight per diet group was recorded for 16 weeks (1 week prior to tumor cell injection). At week 17 of diet treatment, mice were subcutaneously injected with 4×10⁶ MiaPaCa-2 human pancreatic cancer cells (American Type Culture Collection (ATCC), Manassas, VA) in the right flank. Tumor growth was monitored and recorded as previously described for 6 more weeks. Mice were then fasted for 12 hours, anesthetized with isoflurane then killed by cervical dislocation. Tumors were excised and weighed then prepared as previously described. Whole blood was also harvested and prepared as previously described. One mouse from each group did not develop any tumors so were excluded from analysis, thus final sample size is 14.
Prior to injection, the MiaPaCa-2 cells were maintained in DMEM with 10,000 U/mL penicillin, 10,000 U/mL of streptomycin, 1X Non Essential Amino Acids, 10 mM HEPES buffer, and 1 mM sodium pyruvate.

MCF-7 (p53 wild-type) and MDA-MB-231 (p53 mutated) human breast cancer cells and MIA PaCa-2 human pancreatic cancer cells were obtained from the American Type Culture Collection (ATCC) (Manassas, VA).