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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. MiF-6 (TS143344) and M-7 (ATCC CRL-2804) are derivatives of MC-TGS17-51 (ATCC CRL-2799). M7 cells were constructed by transfection of MC-TGS17-51 cells with the pCR3.1 vector alone. The vector contains cytomegalovirus (CMV) and SV40 viral sequences and the neomycin resistance gene. M7 cells are used as a control for MiF-6. MiF-6 cells were constructed by transfection of MC-TGS17-51 cells with pRNAi-5 ligated to pCR3.1 to produce a construct expressing mouse double stranded short-interfering ribonucleic acid (siRNA) formaldehyde dehydrogenase (FDH) mRNA. The pRNAi-5 construct targeted the sequence from -18 to +4 of the mRNA, including the start codon. M7 and MiF-6 cells are 6-thioguanine sensitive, neomycin (G418) resistant, hypoxanthine phosphoribosyl transferase positive (HPRT+) and will grow in HAT selection medium. 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.

male

mouse siRNA formaldehyde dehydrogenase (FDH)

mouse siRNA formaldehyde dehydrogenase (FDH)

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. MiF-6 (TS143344) and M-7 (ATCC CRL-2804) are derivatives of MC-TGS17-51 (ATCC CRL-2799). M7 cells were constructed by transfection of MC-TGS17-51 cells with the pCR3.1 vector alone. The vector contains cytomegalovirus (CMV) and SV40 viral sequences and the neomycin resistance gene. M7 cells are used as a control for MiF-6. MiF-6 cells were constructed by transfection of MC-TGS17-51 cells with pRNAi-5 ligated to pCR3.1 to produce a construct expressing mouse double stranded short-interfering ribonucleic acid (siRNA) formaldehyde dehydrogenase (FDH) mRNA. The pRNAi-5 construct targeted the sequence from -18 to +4 of the mRNA, including the start codon. M7 and MiF-6 cells are 6-thioguanine sensitive, neomycin (G418) resistant, hypoxanthine phosphoribosyl transferase positive (HPRT+) and will grow in HAT selection medium. 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 2 X 10(3) to 6 X 10(3) viable cells/cm2 is recommended.
6. Incubate cultures at 37°C.

Interval: Maintain cultures at a cell concentration between 5 X 10(3) and 3 X 10(5) cells/cm2

Subcultivation Ratio: A subcultivation of 1:4 to 1:6 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