The incubator in the “Dirty” Biobank lab is now clean and running well, so we were able to collect and thaw my tumour baby cells from cryostorage. But first prep…
We are still waiting on our DMEM (liquid medium) shipment so Jo-Maree made up some old (expired) DMEM media with 10% FBS and 1% antibiotics for us to use in the interim.
While it is not ideal, it will enable us to move forward with the project while we wait for delivery and final sign off from the IBC to undertake immortalisation and iPSC protocols.
To ensure we were ready for the cells, we prepped the laminar flow cabinet and T75 flask and warmed 30mL of media in the metal water bath ready to plate the cells when we return from the cyrostores.
The cryostorage area is in the basement of the lab. It features a few large liquid nitrogen cell stores and even has a (rather noisy) liquid nitrogen generator. Access is restricted and everyone needs a buddy when accessing their stores to ensure user safety.
The ‘collector’ (i.e. Jo-Maree) has to rug up with safety gear including nitrile gloves, cryogloves, lab coat, apron and head gear. This protects the user from liquid nitrogen splash/exposure as it can cause severe cold burns.
Jo-Maree stored my three precious fibroid cell vials in rack 12. I am a bit superstitious so see this as a good sign – 12 is my favourite number. They were labelled as PHGL [Post-human Genetic Legacies] 🙂
Jo-Maree picked one of the vials ready for plating up. They need to be kept frozen (on ice) during transport to the lab.
Once we were back in the lab, we added 9mL warm media to the T75 flask, quickly thawed the cells (in my gloved hands with just a sliver of ice remaining) and added them to the flask. During my time at QUT, we often also centrifuged the cells and resuspended them in media to remove any traces of the freezer solution with DMSO. Jo-Maree suggested that this is not necessary in this instance as the DMSO ratio was pretty low.
It will take a few days to see how the cells recover and to check for signs of contamination. It feels good to have them thawed. Fingers crossed for healthy cells….
