Getting started in cell culture work can be an intimidating task. Without all the necessary reagents, or if you use reagents not intended for cell culture use, your culture may encounter problems or fail all together. In this blog, we’re going to cover the important reagents that laboratories may not know they need to have before they get started with a cell culture project.
One if the first things researchers looking to start a cell culture workflow will need to source is serum. Most researchers treat FBS as a standard input, but serum composition can significantly influence experimental outcomes, sometimes masking or confounding treatment effects. If you start your culture and are seeing unexpected signaling, assay variability, or inconsistent drug response, that may indicate the need to evaluate alternative serum conditions such as heat inactivated FBS when complement activity is a concern, human AB serum for immune cell applications, or specialty serum products depending on your assay requirements.
Once you have sourced your serum for your workflow, it’s time to look at Trypsin. Trypsin is widely used for cell detachment, but prolonged exposure or lack of proper neutralization can cleave surface proteins and impact downstream assays, especially flow cytometry and receptor-based studies. Choosing the wrong detachment method can negatively impact CD markers and immune receptors, recovery, cellular viability, experimental variability, and downstream assay accuracy. Some options for detachment methods include Trypsin Inhibitors, Collagenases, Serum, and Proteases.
Growth factor and cytokine supplementation can sometimes be overlooked, particularly in primary or immune cell cultures. This can result in poor growth, inconsistent differentiation, or failed cultures. Depending on your cell type and application, you may use samples like IL-2, IL-7, IL-15, TNF-α, IL-17A, and TSLP, among others. Innovative Research carries all these samples, manufactured with cell culture applications in mind.
After you get your growth factors and cytokine supplements in place, it’s time to consider coating reagents. Many primary and stem cells require extracellular matrix components for proper attachment and function. Poor attachment or altered morphology is often a surface issue, not a cell issue. Some examples of coating reagents include collagen I / IV, fibronectin, laminin, poly-L-lysine, and Matrigel.
Next, you should think about sourcing buffer quality & endotoxin-free reagents. Even low levels of endotoxin (LPS) can activate immune signaling pathways, for example TLR4, introducing background cytokine noise or false positives in sensitive assays. You want to make sure that you’re choosing reagents that are labeled Low Endotoxin for your culture workflow. Remember that other factors, such as anticoagulant choice, can significantly impact cell behavior, signaling, and downstream assay performance. It’s important to keep in mind that sample quality begins before cell culture starts.
Some of the last few core components to consider when preparing your cell culture workflow are Cytokine Stability and your Storage Conditions. Things like repeated freeze-thaw cycles, lack of carrier proteins such as BSA, or improper aliquoting can reduce cytokine activity through denaturation or adsorption loss, leading to inconsistent stimulation. If you purchase your reagents through Innovative Research, we offer custom aliquoting services to help you avoid any issues related to your products and storage. To browse all our cell culture related products, visit the Innovative Research website and check out our collection of Cell Culture Products.
Now, with the information covered here, you should be ready to start your own cell culture workflow. If you’re still sourcing reagents, browse our catalog and check out our collection of cell culture products. If you’re just getting started and still have some questions, reach out to our support team and we will happily guide you in the right direction. We can’t wait to help you make that next breakthrough.
