AtT-20/D16vk-F2 cell line小鼠垂体瘤细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心
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- 货 号：AtT-20/D16vk-F2
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AtT-20/D16vk-F2 cell line小鼠垂体瘤细胞株
Product categoryAnimal cellsOrganismMus musculus, mouseClassificationEukaryota, Animalia, Metazoa, Chordata, Vertebrata, TetrapodMorphologysmall rounded cellsTissuePituitary gland; AnteriorDiseaseTumorApplications3D cell cultureSpecific applicationsThis clone has been used successfully by Moore et al. for several DNA mediated transfection studies relating to endocrine and exocrine secretory pathways.The cells are readily transfected using a standard calcium phosphate protocol.CharacteristicsGrowth propertiesLoosely adherentStrainLAF1Genes expressedadrenocorticotropic hormone (ACTH)CommentsThe F2 subclone of AtT-20 (see BioVector-CCL-89) was developed by B. Gumbiner.Tested and found negative for ectromelia virus (mousepox).Handling informationUnpacking and storage instructionsCheck all containers for leakage or breakage.Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.Complete mediumThe 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%.Temperature37°CAtmosphere95% Air, 5% CO2Handling procedureTo insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a new culture flask. .Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product. The cells will slough if the pH of the medium becomes too acidic (lower than 7.5).Subculturing procedureCells grow in patches and pile up. Cultures do not become confluent.Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.Remove and discard culture medium.Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 minutes).Note: Extreme care must be taken to prevent the cells from clumping. Carefully follow the subcultivation protocol .do not over trypsinize. The cells will slough if the pH of the medium becomes too acidic (lower than 7.5).Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommendedMedium Renewal: Three times weeklyReagents for cryopreservationComplete growth medium supplemented with 5% (v/v) DMSO
Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net
Product categoryAnimal cellsOrganismMus musculus, mouseClassificationEukaryota, Animalia, Metazoa, Chordata, Vertebrata, TetrapodMorphologysmall rounded cellsTissuePituitary gland; AnteriorDiseaseTumorApplications3D cell cultureSpecific applicationsThis clone has been used successfully by Moore et al. for several DNA mediated transfection studies relating to endocrine and exocrine secretory pathways.The cells are readily transfected using a standard calcium phosphate protocol.CharacteristicsGrowth propertiesLoosely adherentStrainLAF1Genes expressedadrenocorticotropic hormone (ACTH)CommentsThe F2 subclone of AtT-20 (see BioVector-CCL-89) was developed by B. Gumbiner.Tested and found negative for ectromelia virus (mousepox).Handling informationUnpacking and storage instructionsCheck all containers for leakage or breakage.Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below _x001f_-130°C, preferably in liquid nitrogen vapor, until ready for use.Complete mediumThe 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%.Temperature37°CAtmosphere95% Air, 5% CO2Handling procedureTo insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a new culture flask. .Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product. The cells will slough if the pH of the medium becomes too acidic (lower than 7.5).Subculturing procedureCells grow in patches and pile up. Cultures do not become confluent.Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.Remove and discard culture medium.Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 minutes).Note: Extreme care must be taken to prevent the cells from clumping. Carefully follow the subcultivation protocol .do not over trypsinize. The cells will slough if the pH of the medium becomes too acidic (lower than 7.5).Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.Incubate cultures at 37°C.Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommendedMedium Renewal: Three times weeklyReagents for cryopreservationComplete growth medium supplemented with 5% (v/v) DMSO
Supplier来源:BioVector NTCC Inc.
TEL电话:400-800-2947
Website网址: http://www.biovector.net
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