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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.
May serve as a platform for the evaluation of several chemotherapeutic strategies to treat chordoma as it accurately recapitulates chordoma. The Brachyury expression may facilitate understanding of its role in other epithelial-derived cancers.
The Chordoma Foundation may be able to offer financial assistance for the purchase of this cell line. Please contact [email protected] for more information.
Fresh surgical chordoma tissue was minced in 0.25% trypsin then placed on shaker at 37°C for 30 minutes. The tissue was then triturated and passed through 40 µm nylon mesh. Single cells were plated and the culture expanded. (source: PubMed: 21699479)
CRL-3267 is a primary chordoma cell line from a sacral tumor that expresses Brachyury. This cell line’s classical phenotypic features, such as characteristic physaliferous cells and stable expression of Brachyury, are maintained through serial passaging.
Expresses embryonic transcription factor Brachyury. This cell line was accessioned with the support of the Chordoma Foundation, a non-profit organization working to improve the lives of chordoma patients by accelerating research to develop effective treatments for the chordoma disease.
Thaw ampoule in 37˚C water bath for approximately 2 minutes. Transfer thawed cell suspension to a 15.0 mL centrifuge tube containing 9 mL complete medium. Mix suspension by gentle inversion. Remove 0.5-1.0 mL for cell count. Centrifuge remaining suspension in the 15mL centrifuge tube at 175-195 x g (1000rpm in an IEC HN SII centrifuge or equivalent) for 5 minutes, RT˚. Discard supernatant and gently resuspended pellet in 5ml fresh complete medium. Transfer 5 ml re-suspended cells into 1 T-75 flask containing 10ml fresh medium. Place the cells in a 5% CO2 incubator @ 37˚C
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subcultivation Ratio: 1:2 to 1:3 is recommended.
Medium Renewal: 2 to 3 times a week
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