It’s time to choose a cell line for your project. Research has led you to a few candidates, but narrowing them down can be difficult and the decision goes beyond what works for your experiment. Genetics, identification, sources, reproducibility and a whole lot more are critical characteristics to consider. To help simplify this not so simple process, I’ll break it down into a few topics that you should consider when choosing or working with a particular cell line.
The Basics for Choosing Your Cell Line:
The immediate criteria you consider are: How easy is the cell line to work with? How fast will it grow? And how accessible is it? But next on the list of things to consider, which is critical, is making sure the cell line you choose best mimics the traits you’re trying to study. Does it grow fast or slow? Does it spread easy or not? What do the publications say about each cell line of interest, and which one best represents the work you’re trying to do?
Going Slightly Beyond the Basics:
Your motives are also going to play a role in the cell line you choose. For example, if you’re doing stem cell research, you will need to also consider which cells have a higher differentiation potential for the ectoderm, endoderm or mesoderm. And that will further depend on which cells you want to study. If you want to study kidney cells, you would need to select cell lines that have a higher differentiation potential for the mesoderm.
The Complex Aspects of Choosing Your Cell Line:
When doing very specific research, deciding on your cell line is going to require far more variables. If you’re testing the behavior of a drug, selecting a cell line cannot be as simple as saying “Well I’m studying prostate cancer, so I’ll stick with the classic PC-3 line.” According to Aaron Krol’s article “The New Cancer Models, Part 3: Cell Line Critics,” a cell line that has been repeatedly published is often chosen without other more important considerations such as how closely it falls in line with the genetics of the cancer in question. If approaching cell line selection based on what has been popularly published, without doing rigorous research, you might not get the most accurate picture. Instead, you should take more care in examining your choice behind a particular cell line.
Memorial Sloan-Kettering Cancer Center has developed the cBioPortal, which can help make this task a little easier by providing researchers with a way to visualize and analyze cancer genomics data sets. Another program creating user-friendly benefits is Broad Institute’s and Novartis’s Cancer Cell Line Encyclopedia. The CCLE provides genetic characterization of human cancer models. Krol’s article explained that researchers can use the CCLE to measure cell line information against tumor genetic information and essentially score how closely related the cancer is to the potential cell line.
The Misidentification of Cell Lines:
Combing through endless genetic information may seem very tedious, and the scrutiny can’t stop there. When choosing a cell line and planning your purchase, you should evaluate the history of the cell line you’re interested in. Reproducibility (something GoldBio believes in and has written about) is becoming a trending topic within the scientific community, which makes this step necessary; however, it is also very important to be sure of what you’re using because of the impact your research may have on the public. When you’ve identified the cell line you want to work with, it is not always good practice to obtain it from a neighboring lab, the reason being that you have to spend extra time being certain of what you were given. All too often, cell lines are mislabeled. Moreover, you also need to confirm that there has been no contamination, which can happen in any lab. When it comes to very sensitive research, be sure of the company, quality and source of your cell line. Too many cell lines have been misidentified so it’s important to know that a specific, cited cell line has conformed to its same function over the ages.
The International Cell Line Authentication Committee has a database listing cell lines that are known to be cross-contaminated. While this is a helpful tool for identifying cell lines in question, practicing some quality controls within your own lab while working with your cell line is also necessary.
Beyond Choosing a Cell Line:
There are other questions you might have considered when choosing your cell line. For example, if you’re working with a human cell line cell, will your murine growth factors be compatible? Thankfully, if you shop GoldBio’s wide selection of growth factors, that information will be provided for you, either directly in the product description or in the associated documents, and if you can’t find it, you can always ask. Let’s look at FGF1, Murine. Within the body of the description there is mention that “FGF1 exhibits considerable cross-reactivity…” Furthermore, there is a table below the main description that shows the cross-species homology. You can be assured that flexibility does exist in this example, but this shows you how to double check.
What about storing and reviving your cell line? Animal cell lines can be preserved in either cryogenic freezers or in liquid nitrogen. Due to the expense of cell lines, it is important to be careful when preserving your supply. During the freezing process, temperatures must be lowered slowly. However, when reviving your cells, the thawing process must be rapid. And if you’re working with a more unexplored cell line, you should record the percentage of viable cells using Trypan Blue stain.
When all is said and done, you also want to ensure reproducibility. Even when you have researched the history of your cell line and its genetics, and you purchased your cells from a trusted repository there are more steps to building on and validating your work. While a cell line gives you a great picture of in vitro situations, it’s important to then research primary cells to use in order to test your experiment. Primary cells are not ideal for initial research because they hold some significant limitations such as being slow to grow, having limited life spans and little to compare with. However, they offer the benefit of giving you a better in vivo picture. Therefore, it’s good practice to replicate the results of your experiment in primary cells.
The takeaway from this article is to be very critical when selecting the right cells. Ask the right question, spend time doing the research, purchase all products from trusted providers (don’t borrow anything for sensitive experiments), be able to replicate and be sure your lab follows the same procedures. Even though this process isn’t standardize, there are some guides available, and there is nothing wrong with developing a standard for your lab based on published resources.
Broad Institute, Novartis Institutes for Biomedical Research, Genomics Institute of the Novartis Research Foundation (2013). Broad-Novartis Cancer Cell Line Encyclopedia (CCLE). Retrieved August 10, 2015
Gao et al. Sci. Signal. (2013) & Cerami et al. Cancer Discov. (2012). cBioPortal. MSKCC. Retrieved August 10, 2015.
International Cell Line Authentication Committee (October 10, 2014). Database of Cross-contaminated or Misidentified Cell Lines. Version 7.2. Retrieved August 10, 2015.
Krol, A. (2013). The New Cancer Models, Part 3: The Cell Line Critics. Bio-IT World. Retrieved August 10, 2015.
Yu, M., Selvaraj, S., Liang-Chu, M., Aghajani, S., Busse, M., Yuan, J., . . . Neve, R. (2015). A resource for cell line authentication, annotation and quality control. Nature, 520(7547), 307-311. doi:10.1038/nature14397
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