Maximize Your Time in the Lab: Improve Experimental Reproducibility with Thaw-and-Use Cells

Many cell biology researchers can name their department’s  or institutions’s “cell culture wizard”—the technician with 20+ years of experience whose cell cultures are always free from contamination, exhibit reliable doubling rates and show no phenotype or genotype weirdness. Cell culture takes skill and experience. Primary cell culture can be even more difficult still, and many research and pharmaceutical applications require primary cells.

Yet, among the many causes of failure to replicate study results, variability in cell culture stands out (1). Add to the normal challenges of cell culture a pandemic that shut down cell culture facilities and still limits when and how often researchers can monitor their cell culture lines, and the problem of cell culture variability is magnified further.

Treating Cells as Reagents

A good way to reduce variability in cell-based studies is to use the thaw-and-use frozen stock approach. This involves freezing a large batch of “stock” cells, then performing quality control tests to ensure they respond appropriately to treatment. Then whenever you need to perform an assay, just thaw another vial of cells from that batch and begin your assay—just like an assay reagent! This approach eliminates the need to grow your cells to a specific stage, which could take days and introduce more variability.

Continue reading “Maximize Your Time in the Lab: Improve Experimental Reproducibility with Thaw-and-Use Cells”

3D Cell Culture Models: Challenges for Cell-Based Assays

3D Cell Culture Spheroid
3D Cell Culture Spheroid

In 3D cell culture models, cells are grown under conditions that allow the formation of multicellular spheroids or microtissues. Instead of growing in a monolayer on a plate surface, cells in 3D culture grow within a support matrix that allows them to interact with each other, forming cell:cell connections and creating an environment that mimics the situation in the body more closely than traditional 2D systems. Although 3D cultures are designed to offer a more physiologically accurate environment, the added complexity of that environment can also present challenges to experimental design when performing cell-based assays. For example, it can be a challenge for assay reagents to penetrate to the center of larger microtissues and for lytic assays to disrupt all cells within the 3D system.

Earlier this week Terry Riss, a Senior Product Specialist at Promega, presented a Webinar on the challenges of performing cell-based assays on microtissues in 3D cell culture. During the Webinar, Terry gave an overview of the different methods available for 3D cell culture, providing a description of the advantages of each. He then went on to discuss considerations for designing and optimizing cell-based  assays for use in 3D culture systems, providing several  recommendations to keep in mind when performing cell viability assays on larger microtissue samples. Continue reading “3D Cell Culture Models: Challenges for Cell-Based Assays”

Considerations for Successful Cell Based Assays II: Cell Culture Conditions

This is the second in a series of blog posts covering topics to consider when designing and performing cell-based assays. In the first installment, we discussed the importance of choosing the right cell type for your assay. Here we will discuss how cell culture conditions can influence assay results. Continue reading “Considerations for Successful Cell Based Assays II: Cell Culture Conditions”