I am working with MCF7 breast cancer cell lines. Now I want to shift my lab. I am culturing the cells and maintaining in cell culture flask. Now how can i transfer this cell lines to another lab. How can I do it keeping the cells alive?
For transportation of cells, split cells and fill the media up to the brim of culture flask. Make sure that you are using flasks without filters in their caps. Pack the flask properly to protect cells from shocks during transportation. Maintain cells at a temperature of around 30-35 C and make sure that cells reach the destination within a day or so. We receive cells in this condition from our national cell repository. Cryopreserve cells for their long term storage. As mentioned by Robert freeze cells in a freeze medium. Initial freezing is slow and is done in isopropyl alcohol container to achieve a cooling rate of 1 oC/min. This is done to avoid formation of ice crystals. Keep cells at -80 oC for 2-3 days. Transfer cells to liquid nitrogen storage tank for prolonged storage. Do not keep cells for prolonged time at -80 oC as it results in loss of viability of cells. Cryopreserved cells can also be transported. On the day of transportation, transfer cells to dry ice and thaw cells immediately on arrival. We receive cells in this way from commercial sources. Cells remain healthy in both conditions provided that the time of transportation is less.
do you mean you wan to transfer them while having them in flask? I dont think it would be efficiently done as you will have all your cells dead. as I can imagine they are so sensitive to the environment changes. the best way is to freeze and put them into the nitrogen and then relocate them.
Transferring the cells while in flask is not safe as it can lead to contamination or death of cells.
Trypsinize the cells from flask and freeze them in cryotubes, using freezing media. Store them in -140C freezers. While shifting them, transfer them into ice box and thaw in your new lab. Immediately culture them in new flasks. Passage them atleast once, before utilizing the cells for any assays.
Agree with above. For storage, I trypsinize my cells and re-suspend the cell pellet in pure FBS+10% DMSO, before transferring to a freezing block at -80oC. To recover, just defrost an aliquot in the water bath for a few minutes, then add back to a flask with warmed cell culture medium.
For transportation of cells, split cells and fill the media up to the brim of culture flask. Make sure that you are using flasks without filters in their caps. Pack the flask properly to protect cells from shocks during transportation. Maintain cells at a temperature of around 30-35 C and make sure that cells reach the destination within a day or so. We receive cells in this condition from our national cell repository. Cryopreserve cells for their long term storage. As mentioned by Robert freeze cells in a freeze medium. Initial freezing is slow and is done in isopropyl alcohol container to achieve a cooling rate of 1 oC/min. This is done to avoid formation of ice crystals. Keep cells at -80 oC for 2-3 days. Transfer cells to liquid nitrogen storage tank for prolonged storage. Do not keep cells for prolonged time at -80 oC as it results in loss of viability of cells. Cryopreserved cells can also be transported. On the day of transportation, transfer cells to dry ice and thaw cells immediately on arrival. We receive cells in this way from commercial sources. Cells remain healthy in both conditions provided that the time of transportation is less.
I think you should do all of the suggestions above. Freeze cells, store at -80 and liq. N2 and transfer them live. Doing all these methods will increase your chances of getting cells there alive.
Thank you all for your good suggestions. Actually I am very new in cell culture works.
@ Sood- Could you please clarify what you meant my splitting the cells. ow to do that?
There are cryoshippers available...So you can transport your cells in the vapour phase of liq.N2.The dryshippers need to be charged and then your frozen cells transferred into the dryshippers and then relocated to another lab.
Splitting the cells means you treat cells with trypsin-EDTA solution and seed cells in new TC flasks. After each split passage number of cells is incremented by 1. Splitting cells and keeping them for a day or so will ensure that cells are healthy and are in logarithmic phase during the transportation.
Dear Charan,
If its just transferring from your lab to a neighboring lab, just take the flask as it is..nothing will happen. either parafilm the flask or wrap it in aluminium foil to avoid contamination.
If u want to transfer a longer distance, like another lab in another place in the same city, etc. just grow the cells in a T25 flask, when they are around 70-80% confluent fill the medium up to the brim and transfer (dont forget to parafilm the flask).
If u want to transfer for very longer distances, like between cities, just trypsinize the cells when they are 70-80 % confluent and resuspend them in FBS with cryoprotectant, freeze then to -80 atleast and transfer on dry ice.
As for the freezing of the cells, just tryipsinize them again when 70-80% confluent and resuspend them in FBS+10%DMSO in a cryovial ( you can also put them in growth medium with 10%FBS and 10%DMSO also) and put them in an isopropanol box for overnight in a -80 freezer. isopropanol box, lets the vial cool slowly and gradually. next day put the vials in a box in liquid nitrogen.
In case u dont have an isopropanol; box, just put the cryovials at 4 degrees for 1 hour, then the deep freezer in the fridge for 1 hour, then -20 for 4 hours, then at -80 for overnight. next day put them in liquid nitrogen.
OOPS sorry Mr. Moyeenul. I thought it was Mr Charan who asked the quesion.
Can we Lyophilize (Freeze drying) the cells and transport them??
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