8 Steps to Manage Cell Culture Development and Cell Culture Protocols
Do you dream in ATC’s and G’s? Have a favorite cell line? Are you friends with the consumable vendors? If so, we’ve got what you need in the Thermo Fisher™ Platform for Science™ Marketplace solution.
The new Cell Culture and Development Applications round out your bench work, by including biologics reagent generation sample tracking and process management to our out-of-the-box solutions.
Many of our customers are leveraging Thermo Fisher™ Platform for Science™ software to take their starting material, such as a sequence, to the next step through sample prep, cloning, and into recombinant cell lines. Because we know you need cell culture data management as well, we’ve created ready-to-go configurations of these very processes.
Using Applications for Processes in Cell Culture Development
Let’s put take a look at the following scenario.
You’ve been tasked with creating a cell line that overexpresses a gene. This cell line needs to be biologically relevant, have low background for variant proteins, and your target of interest needs to be present on the surface for use in immunology studies. Where do you start?
Step One: Determine Your Insert Sequence
You know your gene of interest. Is it available in house? Do you need to have it synthesized?
A quick query for your target name, gene alias, or DNA sequence, will let you know if this material is available. Check your inventory to see where it is located and how much is left.
Check to see if you have variations on this sequence that might be of use…do you have a tagged version, a sequence with a signal peptide? This information, captured in Platform for Science software, might help you determine the best sequence to use for your intended purpose.
Step Two: Determine Your Vector Backbone
Check out your registered vectors to see what you have and how to use it. Download product sheets, links to websites, and view sequence features to determine the best locations to drop in your insert sequence.
Step Three: Prep Your Samples and Clone
Prep your insert and vector leveraging PCR and restriction digests. QC your results on a gel and purify your sample before you ligate.
Ligate your sequence and vector to create your new construct, transform into bacteria, clone on agar, pick colonies, purify DNA, and screen by PCR.
All of these processes are recorded in the platform using our Molecular Biology application. Event reaction conditions, temperatures, times, gel images, and more, are recorded and stored, allowing you to check your work and share your information with colleagues.
(You can even QC your samples by NGS and track library prep and sequencing steps through the use of NGS applications.)
Step Four: Find Your Cell Line
Since the platform has the ability to track any sample types, fields, and relationships for reference, it is easy for you to record information about an unlimited number of mammalian cell lines (and insect, primary, iPSCs, and stem cells, if you need it).
Perform your search to find your parental cells lines – will you choose HEK, CHO, U2OS, SY5Y? The possibilities are only limited by your business needs.
Step Five: Check Your Inventory
From cryovials, to t-flasks, shake flasks, large vessels like wave bags, and everything in-between, your frozen cells and live cultures are inventoried in the platform. Track number of cells per container, total volume, and exact location in incubators or freezers.
Step Six: Thaw or Passage Your Cells
At this point you’ve identified your cell line and you know its availability.
Grab that cryovial or live cell culture and use Platform for Science software to track viability, cell counts, volume in a culture vessel, media recipes, vessel types dissociation buffers, and more.
Record media changes as you get your cells ready for transfection.
Step Seven: Create Your New Recombinant Cell Line
Transfect! (Or transduce, if you have a viral delivery system you’d like to use.)
Track the process and conditions used for transfection of your new construct into your cell line of choice.
Track when you add your selective reagent, record your new media recipe.
When the time is right, cryopreserve your pool and use the platform to create and track the storage of your new cryovials and annotate your cell line to denote biosafety levels, recombinant status, storage conditions, and more.
Step Eight: Clone Your Cells
Marketplace applications give you a choice – traditional dilution cloning, cloning in semi-solid media, or a little bit of both.
Again, enter information about your conditions, cell counts, viability, media, and more. Record container transfers from vessels to plates, including 6-well plates for semi-solid cloning, and 96-well for dilution cloning and clone picking.
Add additional configurations to the platform to process new clones through FACS, Western Blots, ELISAs, etc.
A Platform to Track Your Lab Material from Start to Finish
Platform for Science software tracks your material’s every move. From initial material, to primers used in PCR, restriction enzymes used to prep the vector or QC the construct, from transfections and new cell lines, through cloning and more. At any point in this process you can review the steps you’ve taken and how far along you are in creating that new cell line.
Use the information tracked through experimental processes, to determine next steps or any necessary construct or cell line redesign.
Automatically forge relationships in the platform, allowing you to look back and ask where your material was used, what parents and children have been used or created, and what your inventory has available.
And this is only the beginning. Use these applications in conjunction with screening apps, in vivo studies apps, NGS apps, or work with us to configure processes to fit any one of your innovative workflows.