My newly reprogrammed iPSC colonies were growing extremely slowly. I tested it for Mycoplasma and it came back positive. Since these are precious cells reprogrammed from patient blood, I want to save them. Please suggest efficient methods for treating these lines for mycoplasma. Thanks.
Invivogen Plasmocin can help you with it.
https://www.invivogen.com/plasmocin
But be ready that:
a) it could be tough and long process
b) you need to control phenotype of your iPSC because treatment can trigger differentiation
c) you need do regular cleaning of BSC, incubator and instruments from mycoplasma; especially pipettes. And keep your aseptic technique perfect
d) better start multiple samples with different treatment concentrations and then select the best preserved
Additionally can recommend you Invivogen mycoplasma test strips for fast everyday checks on mycoplasma. Once they showed no mycoplasma you can use more sensitive method to cross-confirm.
https://www.invivogen.com/mycostrip
P.S. I think Invivogen should pay me commission for such good advertisment :-D
:D Thanks a lot for your detailed reply. Also, when treating iPSCs, do you have to make a single cell suspension or can you treat the colonies as such?
Rituparna Chaudhuri for treatment you do regular cell passaging/media change. Just add Plasmocin to culture media instead antibiotic-antimycotic.
Don't do single cell passaging as it together with Plasmocin can trigger lost of iPS phenotype. You can do single cell passaging later after treatment if you want to do isolation of best colonies.
Hi,
You have a couple of commercially available options that are commonly used for iPSC cultures:
In both cases, follow the manufacturer’s treatment cycles precisely and confirm clearance with at least two consecutive negative tests before expanding or banking the rescued line.
Good luck!
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