Live cell transportation - (Dec/14/2006 )
I might have to agree with this. You know what, I split the original T75 flask received to 8 T75 flasks with antibiotics in the medium for banking and a T25 - for mycoplasma testing - without antibiotics. The next day I got fungal contamination in T25 and the original T75 which I refed - just as a backup (with medium containing antibiotics). But the 8 T75 flasks that I split are good - absolutely no contamination at all!
So yes there is risk in transporting live cells. But since so many of you confirmed that it is regular practice to transport live cells that way, maybe it should be fine. As far as contamination is concerned - many many possible explanations right?
Thank you everybody.
Dear Scifi,
Having bought and handled cells for 30 years, the vast majority of my collegues over the years buy frozen cells.
All cellular academic papers that I have read always have in their material and methods sections "cells were obtained from the ATCC". It would be very interesting to find out if they were delivered Frozen or live. My experience with cells of all kinds is that many perimeters change cells and therefore have to be finely controlled :-
Temperature
pH
Growth media +co factors (or the lack of them)
CO2
FCS
TC Plastics
Mycoplasma/Bacterial Contamination
If any of these vary then this can and will lead to variability or may even cause the cells to change biochemically/pharmacologically. I can see no advantage in buying live cells. If my ATCC frozen cells arrive thawed I always get a replacement vial. The frozen cells take a few days to recover but while this is taking place there is always other things to do in the lab. Frozen storage of cells is also very important. There was a thread a while ago about storing cells at -80oC. Long term storage of cells is always best at -196oC (liquid nitrogen) and this has been proven for years. Why do people want to reinvent the wheel.
Scifi,
Shipment of live cells is a long standing and common practice. I've ordered live cultures from ATCC in the past, although it has been a number of years since I ordered them this way. Most of the cell lines I currently order are cryopreserved. The main reason I believe is you can cryo the cells and keep low passage numbers on Cell Banks and you're able minimize labor by not having to perpetually maintain cultures.
Here are my keys to shipping live cells:
1. Fill the flask to the brim with fresh medium. The main reason for this is to prevent sloshing of the media and damage to the cells. If the cells do detach during shipment they'll normally adhere within a few days of culturing at 37C.
2. Seed the culture at a low density, which will depend on the cell line and its growth characteristics. This is to prevent the cells from either reaching confluency or from metabolizing all of the nutrients in the medium. Lactic acid build-up is a major reason for pH shifting in culture, so the low seed density will minimize this.
I hate the contamination. One thing I've done when shipping live cells is to wrap the cap in parafilm. Keeps the lid tight and minimizes dust and moisture contact on the neck of the flask. I'll frequently wipe the outside of the flask with ethanol as an added precaution when receiving fresh cells in live culture. Just make sure you record the information on the flask before you wipe it down, I've made this mistake.
Today, I received a live flask of mammalian cells from out of state in USA. I immediately put the flask in the incubator. The cells looked fine.
My question is how are live cells in culture transported from out of state location?
I got them by regular overnight FEDEX. Is this correct? Aren't the mammalian cells to be kept in incubator all the time with carbon dioxide? I guess the shipping was atleast for 6 hrs or maybe more! Will the cells survive without incubation for that long? Also the flask was filled with medium to the brim. ~ 210 ml in a T75. I removed the excess medium after ~1hour of incubation. I am very eager to see how the cells will fare tomorrow. But I am just curious, if anyone had experience with shipping/receiving/transporting mammalian cells to other states. We all know how frozen cells are transported - by dry ice right? How about live cells in flasks?
Thanks.
The transportation method you described for live cells is fairly common. We usually get our cells frozen but I have shipped live cells out to our other lab using this method. I usually send a 50-60% confluent flask, filled to the brim with fresh media overnight via fed/ex in a styrofoam box containing room temp ice packs (to keep the flask from moving around... tho im sure you could use peanuts or something else as well). Fed/Ex picks up at 2-3pm here and the cells don't arrive till the following morning or afternoon, so they've been hangin out for about 15-20 hours at r/t in a styrofoam box
I'm not sure what they do with the flask once they get it, I would assume they either remove the media and replace with appropriate amount of fresh or passage immediately.
I might have to agree with this. You know what, I split the original T75 flask received to 8 T75 flasks with antibiotics in the medium for banking and a T25 - for mycoplasma testing - without antibiotics. The next day I got fungal contamination in T25 and the original T75 which I refed - just as a backup (with medium containing antibiotics). But the 8 T75 flasks that I split are good - absolutely no contamination at all!
So yes there is risk in transporting live cells. But since so many of you confirmed that it is regular practice to transport live cells that way, maybe it should be fine. As far as contamination is concerned - many many possible explanations right?
Thank you everybody.
When transporting cells this way, the vents in the cap need to be taped shut with parafilm. Also obviously the flask went around completely contaminated areas (lots of dirt in the cargo handling), so I usually douse the flask liberally with 70% Ethanol before putting it into the flowhood. Flaming the rim after opening also helps.