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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
To insure 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.
Coating description: Dilute rat tail type I collagen (BD Biosciences, Catalog No. 354236) to 50 μg/ml. Add 7.5 ml coating buffer to flask and incubate at room temperature for one hour. Carefully aspirate remaining solution. Rinse flask 2 times to remove acid, using 1x DPBS. Coated flasks may be used immediately or stored at 2-8°C up to one week under sterile conditions.
Volumes used in this subculture protocol are for a 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
D5S818: 11, 12
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Liu X, et al. Establishment and Characterization of a Novel Chordoma Cell Line: CH22. J. Orthip Res. 30 (10): 1666-1673, 2010. PubMed: 22504929