tumor model

The Mutatect mouse tumor model is a series of cell lines (eight are available from ATCC) derived from a murine fibrosarcoma that can grow in culture as well as form subcutaneous tumors in syngeneic C57BL/6 mice. The MC1A fibrosarcoma was induced by methylcholanthrene injection into a C57BL/10 mouse. The resulting tumor was adapted to in vitro growth and a variant named MC1A-C1 was isolated that was capable of growth both in culture and as subcutaneous tumor in 8 to 9 week old female C57BL/6 mice [PubMed 106311342; PubMed: 7577474]. MC1A-C1 cells were treated with N-methyl-N-nitrosourea (MNU) and selected with low concentrations of 6-thioguanine. 6-thioguanine (6-TG) resistant (HPRT-) clones were isolated and screened for tumorigenicity; clone MC-TRG17 was selected. A spontaneously arising 6-thioguanine sensitive, Hypoxanthine - Aminopterin - Thymidine (HAT) resistant revertant named MC-TGS17-51 (ATCC CRL-2799) was identified by selection in HAT. The cells are hypoxanthine phosphoribosyl transferase positive (HPRT+) [PubMed 106311342; PubMed: 7577474]. The Mutatect cell lines can be used in vitro/in vivo for the detection of deletion mutations of the hypoxanthine phosphoribosyl transferase (HPRT) gene. They are also useful for the study of inflammatory infiltration of solid tumors.

The resulting tumor was adapted to in vitro growth and a variant named MC1A-C1 was isolated that was capable of growth both in culture and as subcutaneous tumor in 8 to 9 week old female C57BL/6 mice [PubMed 106311342; PubMed: 7577474].

male

Yes, forms subcutaneous tumors in syngeneic C57BL/6 mice.

The Mutatect mouse tumor model is a series of cell lines (eight are available from ATCC) derived from a murine fibrosarcoma that can grow in culture as well as form subcutaneous tumors in syngeneic C57BL/6 mice. The MC1A fibrosarcoma was induced by methylcholanthrene injection into a C57BL/10 mouse. The resulting tumor was adapted to in vitro growth and a variant named MC1A-C1 was isolated that was capable of growth both in culture and as subcutaneous tumor in 8 to 9 week old female C57BL/6 mice [PubMed 106311342; PubMed: 7577474]. MC1A-C1 cells were treated with N-methyl-N-nitrosourea (MNU) and selected with low concentrations of 6-thioguanine. 6-thioguanine (6-TG) resistant (HPRT-) clones were isolated and screened for tumorigenicity; clone MC-TRG17 was selected. A spontaneously arising 6-thioguanine sensitive, Hypoxanthine - Aminopterin - Thymidine (HAT) resistant revertant named MC-TGS17-51 (ATCC CRL-2799) was identified by selection in HAT. The cells are hypoxanthine phosphoribosyl transferase positive (HPRT+) [PubMed 106311342; PubMed: 7577474]. The Mutatect cell lines can be used in vitro/in vivo for the detection of deletion mutations of the hypoxanthine phosphoribosyl transferase (HPRT) gene. They are also useful for the study of inflammatory infiltration of solid tumors.

Protocol:

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
   An inoculum of 5 X 10(3) to 8 X 10(3) viable cells/cm2 is recommended.
6. Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 1 X 10(4) and 2 X 10(5) cells/cm2

Subcultivation Ratio: A subcultivation of 1:4 to 1:8 is recommended

Medium Renewal: Two to three times weekly

Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C