1. Concept of cell cryopreservation
In the process of scientific research, cryopreservation is a commonly used technology to preserve cells and tissues at extremely low temperatures. The temperature reduction can effectively delay the metabolism, active transport, enzymatic reactions and other life activities at the microscopic scale in the living body.
Placing cells in liquid nitrogen at -196℃ for cryopreservation can temporarily remove the cells from the growth state and preserve their cell characteristics, and then revive them for experiments when needed. In this way, the cells can be revived for experiments when needed. It can prevent the loss of cells due to contamination of the cells being cultured or other unexpected events, and play a role in cell preservation.
2. The significance of cell cryopreservation
-Protect important cell lines and facilitate subsequent experimental operations.
-Preserve human and animal cells for transplantation.
-Preserve cell banks for clinical and research purposes.
3. Principle of cell cryopreservation
Cell cryopreservation is a technique of placing cells in a low-temperature environment to reduce cell metabolism for long-term storage. When cells are frozen without any protective agent, the water inside and outside the cells will quickly form ice crystals, which will trigger a series of adverse reactions:
Some proteins are denatured due to the increase in local electrolyte concentration and the change in pH value, resulting in changes in the internal spatial structure of the cell.
The formation of ice crystals in cells and the denaturation of proteins and enzymes on the cell membrane system will hinder the energy metabolism of the cell.
The lipoprotein complex on the cell membrane is easily damaged during freezing, causing changes in the permeability of the cell membrane and loss of cell contents.
As the freezing temperature decreases, the volume of ice crystals expands, causing damage to the spatial structure of DNA and causing cell death.
Adding a cryoprotectant (penetrating/permeable, CPA) to the solution can protect cells from freezing damage.
[Principle]
The commonly used cryoprotectant is DMSO, which is a permeable protective agent that can quickly penetrate cells, lower the freezing point of cells, inhibit cell metabolism, and slow down cell aging.
Improve the permeability of cell membrane to water, promote the exudation of intracellular water out of cells, reduce the formation of intracellular ice crystals, and thus reduce the damage of ice crystals to cells; when thawing, promote the entry of extracellular water into cells to relieve osmotic swelling. Dilute the electrolytes in the unfrozen solution inside and outside the cells to reduce solute damage.
4. Common methods of cell cryopreservation
In 1972, Dr. Mazur proposed the "two-factor hypothesis" of cell cryopreservation damage, which later formed a theoretical system for cell cryopreservation, and it has become the golden rule for guiding and optimizing cryopreservation for nearly half a century. When the cooling rate is neither high nor low, the cells are moderately dehydrated, a small amount of ice crystals are formed in the cells or no ice crystals are formed, the cumulative effect of "solution damage" and "intracellular ice damage" is minimized, and the cell survival rate is the highest. This cooling rate is called the optimal cooling rate.
In the past few decades, researchers have prepared cryopreservation solutions by mixing culture medium, serum and DMSO in a certain ratio:
The cells are gradually transferred from 4℃, -20℃, -80℃ to liquid nitrogen by hand to achieve the effect of "slow freezing".
The cryopreserved cell suspension is directly placed in a programmed cooling box at -80℃, and then transferred to liquid nitrogen for long-term storage.
Using a programmed cooling instrument, a constant-speed cooling machine with a set program is used to cool the room temperature to -120℃ (below -80℃) at a rate of -1 to -3℃/minute, and then placed in a liquid nitrogen tank for long-term storage.
5. Main ingredients of traditional cell cryopreservation solutions
Dimethyl sulfoxide (DMSO): DMSO is an organic solvent that can reduce the formation of ice crystals when cells are frozen by adjusting the osmotic pressure inside and outside the cells, reducing the damage to cell organs and membranes. DMSO can also inhibit cell metabolism and slow down cell aging.
Fetal bovine serum (FBS): FBS contains rich proteins and growth factors, which can provide nutrition and protection for cells. It can also absorb ice crystals on the cell surface to prevent cells from being damaged by ice crystals.
Heparin: Heparin is an anticoagulant that can prevent cells from agglutinating during cryopreservation. It can also inhibit thrombin on the cell surface to prevent cells from thrombosis.
Glucose: Glucose can provide energy for cells and help cells regain vitality after recovery. It can also inhibit cell apoptosis and protect cells from damage.
6. Advantages of serum-free cryopreservation solution
Serum-free cryopreservation solution does not contain animal serum components. The absence of serum can reduce potential contamination and variation, and provide more stable cryopreservation conditions, which helps maintain the reliability and stability of samples. Ready-to-use products do not require program cooling, are simple to operate, and can be directly placed in an ultra-low temperature refrigerator for storage.
【Product Overview】
Gino Bio serum-free cell cryopreservation solution is a universal cell cryopreservation solution, serum-free, xeno-free, phenol red-free, safe and stable in composition, designed with a combination of permeable cryopreservation protectants and non-permeable cryopreservation protectants, double protection, high cell recovery rate, supports the cryopreservation of common mammalian cells, adult stem cells, induced pluripotent stem cells, primary cells, immune cells, engineered cells, cancer cells and other cells.
【Product Features】
· Safety: serum-free, xenobiotic-free, clear chemical composition
· Efficient: widely applicable to a variety of cells, with good cryopreservation effect on hard-to-freeze cells such as NK
· Stable: simple and clear ingredients, stable product ingredients, stable cell cryopreservation
· Convenient: ready-to-use, supports short-term (6 months) storage of cells at -80°C, supports non-programmed cryopreservation
【Conventional physical and chemical indicators】
· pH neutral: 6.9~7.4
· Osmotic pressure (1:9): 160~190 mOms/kg
· Endotoxin: <0.5 EU/ml
· Mycoplasma: None
· Fungi, bacteria: None
【Product Information】
Part Number | Specification | Storage conditions |
GNMA1042 | 100ml | 2~8℃，18 months |
[Limited trial, apply now]
Trial size: 15ml
How to apply for a trial:
(1) Consult a salesperson or local distributor;
(2) Follow the official account "Genom Bio", reply "Trial", and easily participate in the questionnaire to get high-quality serum-free cell cryopreservation solution immediately!