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Disease
fibrosarcoma
Gender
male
Strain
C57BL/10
Applications
tumor model
Storage Conditions
liquid nitrogen vapor phase
Images
Derivation
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.
Clinical Data
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
Tumorigenic
Yes
Effects
Yes, forms subcutaneous tumors in syngeneic C57BL/6 mice.
Comments
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.
Complete Growth Medium
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing
Protocol:
Remove and discard culture medium.
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.
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.
Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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.
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
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Population Doubling Time
24 hours
Name of Depositor
HC Birnboim
References
Birnboim HC, et al. Mutatect: a mouse tumour model for detecting radiation-induced mutations in vivo. Mutat. Res. 430: 275-280, 1999. PubMed: 10631342
Sandhu JK, et al. Effect of dietary vitamin E on spontaneous or nitric oxide donor-induced mutations in a mouse tumor model. J. Natl. Cancer Inst. 92: 1429-1433, 2000. PubMed: 10974079
Haqqani AS, et al. Expression of interleukin-8 promotes neutrophil infiltration and genetic instability in mutatect tumors. Neoplasia
Haqqani AS, et al. Constitutive expression of interleukin-8 by Mutatect cells markedly affects their tumor biology. Carcinogenesis 22: 243-250, 2001. PubMed: 11181444
Wilkinson D, et al. Hprt mutants in a transplantable murine tumour arise more frequently in vivo than in vitro. Br. J. Cancer 72: 1234-1240, 1995. PubMed: 7577474