Description: MCC6 is a suspension human Merkel cell carcinoma cell line, derived from a secondary tumour of the lymph node of a 70 year old male patient.



Description Key Words: Human Merkel carcinoma



Also Known As: No other names



Organism: Human



Tissue: Metastatic, derived from lumph node



Growth Properties: Suspension culture, slow growing



Morphology: Small cells in tight clusters and balls



Growth Medium: RPMI 1640 (with 2mM L-Glutamine + 25mM HEPES) + 15% Foetal Calf Serum + 10ug/ml Insulin, 5.5 ug/ml Transferrin, 5ng/ml Sodium Selenite (ITS: Sigma Catalogue number I3146-5ML 100x solution)



Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line.



Subculturing Procedure: Seed 1 vial of MCC6 into 1xT25 flask, after 3-4 days it can be transferred to a T75 flask with the addition of 50% fresh medium.

Note that this cell line can be challenging to resuscitate from thaw, is slow growing and favours conditioned medium. This cell line does not grow well when seeded at low density or when clusters are broken down to single cells.



This cell line was originally derived and cultured using a feeder cell line. CellBank Australia has cultured the cell line without the use of a feeder layer but with the addition of insulin, transferrin and selenite (ITS) to the base medium of RPMI1640 and 10%FBS.




Medium Renewal: 1-2 times per week. Feed culture by dilution of 50% fresh medium with 50% conditioned medium where possible. This cell line grows optimally with some conditioned medium.



Subcultivation Ratio: 1:2, split cultures when they reach high density by dilution 1:2. Break up the clusters to smaller clumps by pipetting up and down several times. Do not break the culture down to single cells.



Culture Conditions: Incubate the culture at 37°C with 5% CO2.



Cryoprotectant Medium: 10% DMSO + 90% FCS



Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.

Medium:
Growth Condition:

BioVector NTCC Inc.
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