General Information:
Organism:
Homo sapiens, human
Tissue:
lung
Culture Properties:
adherent
Morphology:
epithelial
Culture Method:
Complete Growth Medium:
RPMI-10(Gibco 31800-022)+10% FBS+Penicillin/Streptomycin
Subculturing:
Volumes are given for a 25cm2flask. 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/v) Trypsin-EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 1.0 to 2.0mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed.Discard Trypsin-EDTA solution.
Add 6.0 to 8.0mL of complete growth medium and aspirate cells by gently pipetting.
Add appropriate aliquots of the cell suspension to new culture vessels.
Cultures can be established between 2×103and 1×104viable cells/cm2. Do not exceed 7×104cells/cm2.
Incubate cultures at 37°C, 5% CO2.
Subcultivation Ratio:
A subcultivation ratio of 1:3to 1:8is recommended
Cryopreservation:
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
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