In vivo tumor models
Subcutaneous tumor models
Appropriate number of cells will be prepared into single cell suspension in 50-100μL volume (PBS:matrigel=1:1) and injected subcutaneously on the flank or the abdominal area of the mice.
Orthotopic tumor models
Tumor tissue or cells can be implanted into the specific orthotopic sites of the mice.
Xenograft mouse models
Tumors are generated from human tumor cell lines or tissues, then dissected and digested using an enzyme cocktail of DNase I (Sigma 10104159001), trypsin inhibitor (Sigma T9253-5G), and papain (Worthington LS003126) into a single cell suspension. Cells are mixed with matrigel (Corning 356237, non-phenol red, low growth factor) and injected into mice for passaging at 2-5million cells per 50μl SQ. Additional cells can be frozen down for future usage of P1 and P2 passages. This is a useful method for cells that lose proliferative capacity after being cultured in plates.
Mammary fat pad model
A midline incision will be made through skin and fascia. Mammary fat pad will be identified and breast carcinoma cells will be injected at the concentration of 1-5X106 in 30μL PBS or with 50% matrigel. Wounds will be sealed with suture or would clip.
Cells will be labeled with luciferase or other reporter genes. Appropriate number of cells will be prepared into single cell suspension in <50μL of PBS. Cells will be injected through tail vein or retro-orbitally.
Cells are injected into the kidney capsule for teratoma formation. Incision will be made on the back near the kidney sites. About 20-50μL of ~2X106 cells will be injected into the kidney capsule. Muscle incision is sealed with absorbable suture and skin is sealed with wound clip or non-absorbable suture.
Tumor fragments from tumor growing in donor mice will be passaged to recepient mice.
Tumor tissue culture
Tumor cells can be cultured by our staff in preparation for in vivo studies.