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209/mdct cell line细胞株
Product categoryAnimal cellsOrganismMus musculus, mouseCell typedistal convoluted tubule cellsMorphologyepithelial-likeTissuekidneyApplications3D cell cultureProduct formatFrozenStorage conditionsVapor phase of liquid nitrogenGeneralSpecific applicationsUseful for studies of Na transport, implications of Gitelman’s syndrome, NaClcotransporter, ENaC, PTH signaling, calcium transportCharacteristicsGrowth propertiesAdherentAge44 to 55 daysGenderMaleImmortalization methodAdenovirus 12 - SV40 virus hybrid (Ad12-SV40)Expression markersNaCl cotransporter (SLC12a3), expressed; PTH receptor, expressedComments209/MDCT is a continuous line of distal convoluted tubule (DCT) cells expressing a phenotype including NaCl cotransporter (Slc12a3) and PTH receptors. The DCT is a primary site for regulation of blood pressure by controlling NaCl absorption and for calcium homeostasis by parathyroid hormone. This cell line is of considerable use for studies of hormonal and pharmacological regulation of fundamental ion transport processes and their underlying mechanisms.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 a 1:1 mixture of Dulbecco’s Modified Eagle’s Medium with 1 g/L glucose and 1mM sodium pyruvate (Life Technologies, Catalog No. 11885) and Ham’s F-12 Nutrient Mix (Life Technologies, Catalog No. 11765). To make the complete growth medium, add the following component to the base medium:fetal bovine serum to a final concentration of 5%Temperature37°CAtmosphere95% Air, 5% CO2Handling procedureTo ensure 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). It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).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 sheet. Subculturing procedureNote: Subculture before cultures become 100% confluent. Cells can detach from flask when over-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 0.05% (w/v) Trypsin - 0.02% EDTA solution to remove all traces of serum which contains trypsin inhibitor.Add 1.0 to 2.0 mL of 0.05% 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 37.0°C to facilitate dispersal.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 xg 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.Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:20 is recommendedMedium Renewal: 2 to 3 times per weekSeeding Density: 5.0 x 103 to 1.0 x 104 cells/cm2Reagents for cryopreservationComplete growth medium supplemented with 20% (v/v) FBS and 10% (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, mouseCell typedistal convoluted tubule cellsMorphologyepithelial-likeTissuekidneyApplications3D cell cultureProduct formatFrozenStorage conditionsVapor phase of liquid nitrogenGeneralSpecific applicationsUseful for studies of Na transport, implications of Gitelman’s syndrome, NaClcotransporter, ENaC, PTH signaling, calcium transportCharacteristicsGrowth propertiesAdherentAge44 to 55 daysGenderMaleImmortalization methodAdenovirus 12 - SV40 virus hybrid (Ad12-SV40)Expression markersNaCl cotransporter (SLC12a3), expressed; PTH receptor, expressedComments209/MDCT is a continuous line of distal convoluted tubule (DCT) cells expressing a phenotype including NaCl cotransporter (Slc12a3) and PTH receptors. The DCT is a primary site for regulation of blood pressure by controlling NaCl absorption and for calcium homeostasis by parathyroid hormone. This cell line is of considerable use for studies of hormonal and pharmacological regulation of fundamental ion transport processes and their underlying mechanisms.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 a 1:1 mixture of Dulbecco’s Modified Eagle’s Medium with 1 g/L glucose and 1mM sodium pyruvate (Life Technologies, Catalog No. 11885) and Ham’s F-12 Nutrient Mix (Life Technologies, Catalog No. 11765). To make the complete growth medium, add the following component to the base medium:fetal bovine serum to a final concentration of 5%Temperature37°CAtmosphere95% Air, 5% CO2Handling procedureTo ensure 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). It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).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 sheet. Subculturing procedureNote: Subculture before cultures become 100% confluent. Cells can detach from flask when over-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 0.05% (w/v) Trypsin - 0.02% EDTA solution to remove all traces of serum which contains trypsin inhibitor.Add 1.0 to 2.0 mL of 0.05% 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 37.0°C to facilitate dispersal.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 xg 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.Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:20 is recommendedMedium Renewal: 2 to 3 times per weekSeeding Density: 5.0 x 103 to 1.0 x 104 cells/cm2Reagents for cryopreservationComplete growth medium supplemented with 20% (v/v) FBS and 10% (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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