Trypsin is a proteolytic enzyme secreted as trypsinogen by the pancreas, and activated to aid in degradation of food proteins in the proximal intestine. In the life sciences lab, trypsin is often used to dissociate adherent cells from culture plates, flasks, etc. Traditional trypsin solutions must be stored frozen at -20 °C to prevent autodigestion, which can lead to reduced enzyme activity (Figure 1), resulting in ineffective cell detachment. StableCell™ Trypsin solutions are specifically designed to perform cell detachment as standard trypsin solutions do, without the need to aliquot, freeze, and thaw the trypsin. This saves significant time by eliminating time-consuming aliquoting and waiting for trypsin to thaw for cell passaging or experimentation, not to mention freeing space in your freezer. Our StableCell™ Trypsin solutions are recommended for storage at 2-8 °C, however, our studies indicate, with a high degree of confidence, that StableCell™ Trypsin solutions retain >90% of their activity and performance even when stored at 37 °C.
StableCell™ Trypsin has been validated using a variety of cell lines. Our studies have included SKOV-3, DU145, HEK293, 16HBE41o-, HL-1 and MDCK cell lines. We’ve compared detachment and enzyme activity of StableCell™ Trypsin to our standard Trypsin solutions, and found similar performance throughout for each cell line we observed. StableCell™ Trypsin’s unique formulation, however, allows it to retain its enzyme activity over time, at 2-8 °C, whereas our standard trypsin solutions degrade quickly when not kept frozen. See Figure 1 and Figure 2 for details (StableCell™ in purple, standard trypsin in blue).
Figure 1. Enzyme activity measured over time. StableCell™ Trypsin Solution compared to normal, research grade trypsin – both were stored at 2-8 °C. Results show that while normal trypsin activity tapers off over time, StableCell™ Trypsin retains >90% of it's activity.
Figure 2. Average number of cells detached using both our standard trypsin solutions (Product No. T3924, T4049, T4174) compared with StableCell™ Trypsin solutions (Product No. T2601, T2605, T2610).
Our studies conclusively suggest that StableCell™ Trypsin solutions demonstrate identical cell detachment performance when compared with standard trypsin solutions. (Figure 2).
"There are other solutions for cell detachment which similarly allow for storage at 2-8 °C, or even room temperature. It’s nice not having to wait for it to thaw, but what I like most about StableCell™ is that it’s actually trypsin, so I know exactly what I’m putting in with my cells."
Figure 3. StableCell™ Trypsin vs Normal Trypsin
StableCell™ Trypsin Solution may be used to remove adherent cells from a culture surface for passage, cryostorage, or experiments. Cells are most commonly removed from the culture substrate by treatment with trypsin or trypsin - EDTA. StableCell™ Trypsin Solutions can range from 0.05% to 0.5%. The reasons for the range of concentrations are as follows:
Incubating cells with excessive trypsin concentration or for longer than needed will cause cell injury and death by damaging cell membranes. If unsure about the concentration of trypsin to use, start with the lowest expected effective concentration, and increase only if cells do not detach. Lot-to-lot variation in dissociation times can occur and are to be expected since the enzymatic activity of each lot will differ. If trypsin is being solubilized or diluted from a concentrated solution, it is important to use a buffered salt solution that contains no Ca2+ or Mg2+, such as Hank's Balanced Salt Solution, Modified ( Product No. H9394). Adjust the pH of trypsin solution to 7.4–7.6.
Note: The time required to remove cells from the culture surface is dependent on cell type, population density, serum concentration in the growth medium, potency of trypsin, and time since last subculture. Trypsin can cause cellular damage, so exposure should be kept to a minimum.
Our studies show that StableCell™ retains >90% activity for up to 7 weeks at temperatures up to 37 °C.
Many non-freezer solutions for cell detachment are sold as a replacement for trypsin, and are not actual trypsin solutions. StableCell™ is an optimized trypsin solution, so you can be sure of what you are adding to your cells.
We’ve conducted tests on a small variety of cell lines, including commonly used lines, as well as a few cell lines which have been known to be difficult to dissociate. Our evidence suggests StableCell™ Trypsin works well against the same large variety of cell lines as any other trypsin.
No, studies have been done cellular protein modifications using both StableCell and traditional trypsin and no difference was seen between the samples.