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A technology of biological cells and antifreeze, applied in microorganisms, biochemical equipment and methods, preservation of human or animal bodies, etc., can solve problems such as low survival rate, and achieve expansion of research scope, low production cost, and simple preparation method. Effect
Inactive Publication Date: 2008-07-16
NORTHWEST A & F UNIV
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[0008] Aiming at the current low survival rate of mammalian embryos, sperm, cells and organ tissues after freezing, the purpose of the present invention is to provide a biological cell freezing antifreeze agent with simple preparation method, low cost and good freezing preservation effect
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Embodiment 1
[0030] The crude polysaccharide method that embodiment 1 extracts
[0031] The laminaria polysaccharide described in the present invention is a crude polysaccharide extracted through natural laminaria (Laminaria nponica Aresch.) leaching, concentration, protein removal, dialysis to remove impurities, and alcohol precipitation.
[0032] Concrete steps: wash, dry and pulverize the kelp, add 25 times of distilled water after weighing, heat to 90°C, and extract for 5 hours. Vacuum filtration with a Buchner funnel to obtain the extract. The extract was concentrated 5 times using a rotary evaporator. Then add 1 / 3 volume of chloroform:amyl alcohol (4:1) to the concentrated solution, shake for 30 minutes, centrifuge at 4000 rpm for 30 minutes, collect the supernatant, and repeat 4-5 times. Then use a dialysis bag to remove impurities. Precipitate at 4°C for 24 hours with 4 times the volume of 95% ethanol, centrifuge at 4000 rpm, dry the precipitate, and pulverize it.
Embodiment 2
[0034] Weigh 2.16g of glucose and 1.008g of sodium citrate with an electronic analytical balance, dissolve them in 72ml of distilled water, stir them evenly with a magnetic stirrer to prepare a base solution; add 18ml of albumin to the base solution and mix well, add 100,000 IU Prepare liquid I with penicillin; add 10ml glycerin to liquid I to prepare liquid II; add laminarin to liquid II to make the concentration of laminarin 0.025g / 100ml, adjust the pH value to 6.21, then perform filter sterilization and cool to room temperature , placed in a 4°C refrigerator for later use.
Embodiment 3
[0035] Embodiment 3 The 100ml cell freezing antifreeze consists of the following raw materials:
[0036] Laminarin 0.05g, glucose 2.16g, sodium citrate 1.008g, distilled water 78ml, albumin 15ml, glycerin 7ml, penicillin 80,000 units.
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Abstract
The invention discloses a biological cell freezing antifreeze, and 100ml of the cell freezing antifreeze consists of the following raw materials: 0.025 to 0.3g of kelp polysaccharide, 2.0 to 3.5g of glucose, 0.5 to 1.5g of sodium citrate, 60 to 80ml of distilled water, 10 to 20ml of albumin, 5 to 18ml of glycerol and 0.05 to 0.15 million units of penicillin. The preparation method is that: the glucose and the sodium citrate are dissolved in the distilled water to prepare a base liquid; the albumin is added, the mixture is mixed evenly, and the penicillin is added to prepare a I liquid; the glycerol is added in the I liquid to prepare a II liquid; the kelp polysaccharide is added in the II liquid for even mixing; the pH value is adjusted to 6 to 7.5, then the filtration and sterilization are carried out, and the mixture is cooled until reaching the room temperature and then arranged in a refrigerator of 2 to 5 DEG C for standby. The cell freezing antifreeze is applicable to the cryopreservation of mammalian semen, somatic cells, oocytes, early embryos, organs and tissues, in particular to the cryopreservation of the semen of various mammals.
Description
technical field [0001] The invention belongs to the field of ultra-low temperature biology, and relates to a technology for storing biological tissue materials at ultra-low temperature and maintaining their biological activity after thawing, in particular to a freezing antifreeze agent for biological cells; The preparation method of the drug and its application in the cryopreservation of mammalian somatic cells, oocytes, early embryos and organ tissues; especially in the cryopreservation of various mammalian semen. Background technique [0002] Animal organ tissues or cells are specially treated and stored in ultra-low temperature, which inhibits their metabolic activities and preserves them in a relatively static state. Once the temperature rises, it can be revived without losing life activity. The generally accepted theory of the resuscitation process is that during the freezing and thawing process, under the protection of the cryoprotectant, the ice crystallization of in...
Claims
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Application Information
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