Organism:

Homo sapiens, human

Tissue:

skin/epidermis

Culture Properties:

adherent

Morphology:

epithelial

Complete Growth Medium:

DMEM (Gibco 12800-017)+10% FBS+ Penicillin/Streptomycin

Subculturing:

Volumes are given for a 25cm² 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/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.
• Incubate cultures at 37°C, 5% CO₂.

Subc*tion Ratio:

A subc*tion ratio of 1:3 to 1:8 is recommended

Cryopreservation:

Freeze medium: Complete growth medium supplemented with 10% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase