MLE 12 小鼠肺上皮細(xì)胞
MLE 12 (ATCC® CRL-2110™)
Organism Mus musculus, transgenic, mouse, transgenic
Tissue lung
Cell Type epithelial; SV40 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain Papovavirus]
Age 5 month old
Gender female
Strain FVB/N
Storage Conditions liquid nitrogen vapor phase MLE 12 小鼠肺上皮細(xì)胞
Derivation
This line was established in 1992 by Kathryn A. Wikenheiser from pulmonary tumors in a mouse transgenic for the SV40 large T antigen under the control of the promoter region of the human surfactant protein C gene.
Clinical Data
female
5 months
Genes Expressed
lung surfactant proteins B and C (SP-B, SP-C)
Tumorigenic Yes
Effects
Yes, tumors were generated within 6 to 9 months in nude mice
Comments
The cells express the mRNA for large T antigen.
Lung surfactant proteins B and C were detected.
The cells secrete phospholipids in response to phorbol esters and ATP but not in response to forskolin.
MLE 12 小鼠肺上皮細(xì)胞
HITES medium with 2% fetal bovine serum is formulated at the ATCC as follows:
Dulbecco's medium : Ham's F12, 50:50 mix (ATCC 30-2006)
Insulin 0.005 mg/ml
Transferrin 0.01 mg/ml
Sodium selenite 30 nM
Hydrocortisone 10 nM
beta-estradiol 10 nM
HEPES 10 mM
L-glutamine 2 mM (in addition to that in the base medium)
fetal bovine serum 2%
Subculturing MLE 12 小鼠肺上皮細(xì)胞
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
Remove and discard culture medium.
Briefly rinse the cell layer with 0.25% (w
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) 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.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:5 to 1:10 is recommended
Medium Renewal: Twice per week
Cryopreservation MLE 12 小鼠肺上皮細(xì)胞
Freeze medium: Complete growth medium supplemented with 5% (v
) DMSOStorage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5% MLE 12 小鼠肺上皮細(xì)胞
