Advantages of dissociating your cells with Accutase

An alternative solution to trypsin

Accutase® is an enzyme solution used in cell culture techniques to dissociate adherent cells from a culture surface. It’s a mixture of collagenolytic and proteolytic enzymes, mainly composed of a protease called Trypsin-like protease XIV and a neutral protease called thermolysin. Accutase is a ready-to-use, solution that does not contain any mammalian or bacterial derived components. Accutase is often selected as a more gentle alternative to trypsin, another enzyme commonly used to dissociate cells.

Let’s take a closer look at the top four advantages of using Accutase and what makes Accutase a versatile and smart choice as a cell culture dissociation solution.

Gentle cell dissociation

Accutase is known to be much more gentle on cells and is less likely to cause cellular damage compared to trypsin.  Gentle cell dissociation is important for a number of reasons, particularly when working with sensitive or fragile cell types.  Using gentle dissociation methods helps to maintain cell health, function, and morphology, while minimizing cell stress and damage.

Efficient cell dissociation

Despite being gentle, Accutase is also effective at detaching cells from the culture substrate. Accutase acts relatively quickly to dissociate cells from culture surfaces, typically requiring less time than other enzymes for cell detachment. This makes it a popular choice for routine cell passaging and other cell culture applications.

Effective dissociation

Accutase can help reduce cell clumping, which can improve cell survival and lead to more homogeneous cell populations.  And importantly, Accutase is known for preserving cell surface antigens, which are often lost during the use of other dissociation methods. This makes it a valuable tool for experiments where the expression of surface markers is critical, such as in flow cytometry or immunofluorescence analysis.

No need for serum

Accutase can be used without serum, which is particularly useful when working with serum-free media or when serum components interfere with downstream assays.  In addition, the enzymatic action of Accutase does not generally need to be quenched with serum.  Unlike trypsin, Accutase is gentle enough that only dilution with DPBS or cell culture media is required to stop the dissociation activity.  (In cases where enzyme deactivation is required, Soybean Trypsin Inhibitor is an effective serum-free solution compatible with Accutase.)

What types of cells can Accutase be used with?

Accutase is commonly used with a wide range of cell types, and is particularly useful for dissociating delicate cell types, such as pluripotent stem cells and neuronal cells, as well as for dissociating cells that are difficult to detach from surfaces.

Cells that benefit from Accutase:

• Human pluripotent stem cells (hPSCs) – Both human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) are extremely sensitive to harsh dissociation methods, and are vulnerable to cellular damage, unwanted spontaneous differentiation, and apoptosis when dissociated using trypsin.  The more gentle nature of Accutase can help preserve the “stemness”, pluripotency, and viability of the hPSCs in culture.
• Mesenchymal Stem Cells (MSCs) – Accutase can be used to dissociate MSCs from bone marrow or other tissues, while preserving their viability and differentiation potential.
• Adult Stem & Progenitor Cells – Many adult stem cells and differentiation cultures benefit from dissociation with Accutase, including human neural stem cells (NSC), hematopoietic stem cells (HSC), epithelial stem cells, and dental pulp stem cells. 
• Primary Cells – Accutase can be very beneficial to dissociate sensitive or fragile primary cells, preserving their viability, morphology, and function.
• Cell Lines – Accutase can be used to detach many common cell lines for passaging with minimal cell damage, including HeLa, COS-7, and HEK293T cells.
• Tumor Cells – Some tumor cell types can be difficult to dissociate using traditional methods, including trypsinization.  Accutase

The optimal conditions and protocol details for dissociating cells with Accutase can vary depending on the cell type and culture conditions and downstream application.  It’s important to follow the manufacturer’s guidelines and optimize Accutase treatment for specific experiments.

Summary

Accutase is a versatile, easy-to-use, and effective enzyme blend that offers several advantages for cell culture, including gentle and fast cell dissociation, preservation of cell viability and functionality, and compatibility with various cell types. 

Learn More

• Explore the Accutase® product information page
• Connect with Captivate Bio to learn more about you’re your cells can benefit from incorporating Accutase into your experiments
• See what other researchers have to say about Accutase on ResearchGate

Accutase is a registered trademark of of ICT, Inc.