Organism:

Mus musculus, mouse

Tissue:

embryo

Culture Properties:

adherent

Morphology:

fibroblast

Complete Growth Medium:

DMEM (Gibco 12800-017)+10% CS+ 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. Never allow culture to become compley confluent.

• 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:

The recommended inoculum is 2~3×10³cells/cm². Subculture before cultures become 70 to 80% confluent or when cells reach 5 ~ 6×10⁴ cells/cm².

Cryopreservation:

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

Storage temperature: liquid nitrogen vapor phase