Hi everyone i am culturing Hep G 2215 hepatomoma cell lines.My problem is that after 2 -3 day of culture.Small dot dot like stricture seen in flask( But they have not moving like Bactria or they are like yeast bunch).I have Chang my media and all thing also check my hood and incubator.I am washing cells with BPS 1 x which contain PS antibiotics.This problem come only in this cell lines other cells are OK( Huh-7, HepG-2,SNL, HNF).Some help me plz.
I think they are some extragenous material which we unable see by naked eye. So for checking the media u take small aliquot in flask keep them for 48 h at room temperature. If turbidity observed then media is contaminated.
thank seema .But i have check my media( punting media only in t-25 and kept in co2 incubator).I am growing other cells and taking same media.
I’m not sure if it’s the same problem but I had the same small dot like stricture in my LHC-9 media today, similarly the dots have no movements and they don’t look like any bacterial nor yeast contamination i have ever seen. I thought it might be the additives i added to the medium so i checked my other LHC-9 medium that doesn’t contain additive and guess what the same dot like stricture is present ! . I really don’t know what to make out of them but I filtered few ml and incubated them along with unfiltered ones in 37oc 5% CO2 incubator, I also incubated some filtered medium in the fridge to see if the dot like stricture form again. I guess all left know is to wait for Monday and I'll post an update for it .
You still filter sterilized the media and other addative again before composing the complete growth medias. Culture the cells under most sterilized condition. All the best.
hi Dhananjay , are you using the same media for all your cell lines ? after checking my results yesterday i concluded that the problem is the media it self ! which is kinda strange . It is my first time ever to work with LHC-9 medium and i found similar structure in two different bottles. Even though they are used in different occasions i have opened them around the same time which was less than one month ago therefore i think its something to do with their manufacture or storage ( i received them in -20) and i have kept them in the fridge after i defrosted them the first time. Im not sure if that would help but i thought to share what i found.
Thanx Fatema i am using same media for all cell line Except ESD-3 and H-9 they are hESCs cell line.Right now i have stop culturing this cells.
(And start my work after checking Mycoplasma.Although no symptomam of Mycoplasma contamination)
Thanx alot for your valuable response.
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