Hello,
I am working with zebrafish liver (ZF-L) epithelial cell line (ATCC® CRL2643™).
Contamination in flasks were observed in May after transfer of equipment and materials from another laboratory to our cell culture laboratory.
I used bleach, alcohol and formaldehyde to control contamination. The hepa filter of the incubator was changed. I've decontaminated the incubator several times. I have tried different brand distilled water and flask. Many cell vials were thawed by different people at different times for three months. Despite all these efforts, there are no growth and change in morphology of the cells which are adhere to flask.
Has anyone experienced this problem before?
Could you please give me some advice for progressing with my research?
Thank you for your answer in advance.
yes contamination is a problem while doing cell culture experiments. regarding use of flask maybe you can use corning flasks as they are sterilized by e-beam. for cryopreservation some specific solutions may be used for this cell line. for attachment in flasks may be you can use collagen coated flasks or petri dishes. may be prolonged culture storage may lead to alterations or even EMT transitions leading to loss of cell polarity of zebrafish liver epithelial cells. You can also modify the culture media if you can and use fresh medium with strong antibiotics and perfect filtration of cell medium. lastly since liver itself is a lobular structure may be you can study more literature regarding confluent monolayer establishment of zebrafish liver epithelial cell line.
hope this answer could help
Shubhrajyoti Das
I am using corning cell flasks. In the process of cryopreservation, %5 DMSO was used according to ATCC procedure. Pen-Strep (Streptomycin and penicillin solution) was used in complete media. Ham's F12 and DMEM (Dulbecco's Modified Eagle Medium) were filtered with 0.22 uM filters. But I can't filter serums in complete medium because of their viscosity. I don't know how I can sterilize this serum content. I am going to make a survey of collagen-coated flasks and EMT transitions. Thank you so much for your contribition.
Hi Richard Tan Yi Jer
, I appreciate the valuable information and advice you have shared. I am going to use broad spectrum antibiotics and 0.45 uM filters. Thank you so much.- Badges
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