Application of biological buffers HEPES and MOPS in cryopreservation
"Cryopreservation" is the most common process in the biological field. During cell culture, it is sometimes necessary to store cells for subsequent research or for cell therapy, which can preserve more cells for the patient. During freezing, the unstable changes induced by freezing have different effects on cells. If the operation is not careful, it is very likely that the cells will inadvertently necrosis during the freezing process and fail to meet the requirements. Therefore, scientists take this very seriously and have to study different solution formulations for samples from different cell lines to create the best results.
Continuing the previous stage of cell therapy, this stage will introduce the relevant precautions for cell cryopreservation: Since cell preservation in vitro requires extremely high stability and strict living environment requirements, in order to enable cells to be stored for a long time, cryopreservation technology is used for Cells were stored in liquid nitrogen to allow cellular metabolic reactions to enter stationary phase. Thaw again at a time. During the freeze-thaw process, the medium will encounter problems such as crystallization, solute precipitation, polarity changes, pH changes, and concentration changes. Here we will provide solutions for crystallization, concentration changes and pH changes.
To solve such problems, there are currently two methods: slow freezing method and vitrification freezing method. Through slow freezing, water molecules in cells have enough time to be discharged, and cryopreservation agents are added to prevent water molecules inside and outside cells. Form ice crystals that are too large to dehydrate them as completely as possible. Vitrification freezing: adding a higher concentration of cryopreservative to the culture medium, and using the method of rapid freezing, the water in the cells can be quickly replaced in a short time, so that the liquid in the cells becomes a high-viscosity, high-concentration vitrified substance. Reduce ice crystals.
Results of mixing Na-P (20 – 100 mM) with 50 mM HEPES solution; no major acidity changes were observed over the expected concentration range. In the literature, two Na-P concentrations, 5 mM and 50 mM, were used to compare different HEPES concentrations (0–30 mmol L-1) (Fig. 3), compared with samples without Na-P (Fig. 2) . The H2-value is very obvious.
In the mixed results of 20 mM MOPS and 50 mM Na-P, the H2-value increased to (7.49 ± 0.0503) and the acidity change was relatively stable.
To minimize pH changes, the required MES concentrations are approximately 12 mM in 5 mM Na-P and approximately 80 mM in 50 mM Na-P.
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