I'm currently growing OVCAR3 and OVCAR8 cells in T225 flasks. I returned to the lab a couple of months ago after being away for nearly half a year due to COVID-19, and since I've returned I've had problems with my cells growing very sparsely in some areas of my flask but reaching a high density quite quickly in other areas. One of my labmates is having the same issue with different cell lines (I haven't asked the others so not sure if this is happening to everyone or just us) and we have no idea what's causing it. Neither of us were having this problem before our extended break from the lab. Any suggestions would be greatly appreciated!
some times cells need a while in order to re -grow specially if they kept in the deep freeze or in liquid nitrogen> It will take about 2 to 3 passages until they behave as usual. when you split focus in the cell counting and the make the cells more concentrated than normal. for example 2 ml of cell suspension into 8 ml media and pippet them very well when seeding in order for the equal spread of the mono layer. also check the shelf of the incubator as well if its located properly or not .It happens when cleaning the incubator and placing the shelf in the wrong order.
I hope this can help
best wishes
Shilan
Are your incubator shelves level? Sounds like they might not be. Even the tiniest slope will make a big difference.
You might have to adjust the feet on the incubator to level the shelves out. Also, sometimes the shelves can get warped, especially if they've been autoclaved so check for that.
Hello Melanie Seaton,
I totally agree with Govinda Sharma, Sometimes the answer is in the simplest points.
If you've never had this problem before and you're making everything exactly the same, the issue may be related to the consumables and/or reagents that you used. Be sure to double check your material. Maybe there was a blackout during the COVID lockdown and some of your consumables/reagents are good no more because of changes in temperature, humidity, etc. Another possibility might be that the flasks that you're using (or at least that specific batch) might have some issues. I've been using six well plates a lot for the past 2-3 years and i've seen that in some batches cells tend to stick easier in specific regions of the well. In my case, they form two rings on the center of the well. (one smaller than the other one and centered inside the bigger one). I think this trend might be linked to the process manufacturers use to prepare flasks and plates for cell culture. Since every single time i´ve seen this issue, the pattern is exactly the same.
Hope this helps.
If you can give more details or pictures of your cultures maybe we could be more help.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line