An Improved Trypsin Digestion Solution and Its Application

This study aims to provide an improved trypsin digestion solution and explore its advantages and more applications in cell digestion by comparing it with 0.25% trypsin, trypsin-EDTA (0.25%), and citric acid trypsin. The stability, timeliness, and digestion ability changes of the improved trypsin digestion solution under different storage conditions were evaluated by measuring pH values and observing the degree of cell digestion under a microscope. The dispersion effect between cells and the effects on cell survival rate, proliferation rate, and adhesion after overdigesting cells were verified by calculating the intercellular dispersion rate, trypan blue staining, and CCK-8 cell proliferation detection experiments. The results showed that under different storage conditions, the pH value change range of the optimized trypsin digestion solution was the smallest and its digestion ability was the most stable. Compared with 0.25% trypsin, the optimized trypsin digestion solution could significantly increase the intercellular dispersion rate. Compared with trypsin-EDTA (0.25%), when overdigesting cells, the improved trypsin digestion solution could significantly reduce the cell mortality rate and significantly increase proliferation rate, and adhesion ability. These results indicate that the improved trypsin digestion solution has significant improvements in terms of storage timeliness, stability, and digestion ability. Moreover, it has a better dispersion effect between cells and causes less damage to cells, and has significant advantages in nourishing and stabilizing experimental cell lines and promoting rapid adhesion and proliferation growth of cell lines after passage. This also provides a more suitable cell digestion solution and technical support for experimental cell lines requiring continuous passage culture, cell clinical transformation therapy, counting experimental cell lines, and single-cell research.

CSTR: 32200.14.cjcb.2025.01.0009