A kind of cryopreservation liquid and preparation method thereof
A cryopreservation liquid and solution technology, applied in the field of biomedical materials, can solve the problems of affecting the survival rate of cryopreservation objects, the safety of functional expression, the inability to effectively control the growth of ice crystals, and the lack of ice crystal growth, etc., to achieve high cell survival rate and Functional expression stability, excellent spreading performance, and convenient raw material sources
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[0074] The preparation method of the present invention will be described in further detail below with reference to specific examples. It should be understood that the following examples are only for illustrating and explaining the present invention, and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the intended protection scope of the present invention.
[0075] The experimental methods used in the following examples are conventional methods unless otherwise specified; the reagents, materials, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.
[0076] The PVA used in the embodiment of the present invention has a syndiotacticity of 50%-55%, a molecular weight of 13-23 kDa, and a degree of hydrolysis of 98%.
[0077] In the embodiment of the present invention, the poly-L-proline polymerizat...
Embodiment 1
[0079] Example 1: Cryopreservation of oocytes and embryos
[0080] 1. Prepare cryopreservation solution: prepare cryopreservation solution according to the following formula
[0081] Cryopreservation Solution A:
[0082] Each 100mL contains the following components:
[0083] substance content PVA(g) 2.0 Ethylene glycol (mL) 10 DMSO(mL) 10 Sucrose (mol L -1 )
0.5 Fetal bovine serum (mL) 20 DPBS(mL) margin
[0084] 2.0 g of PVA was heated in a water bath at 80°C and dissolved in 25 mL of DPBS with magnetic stirring. After the PVA was completely dissolved and cooled to room temperature, the pH was adjusted to 7.0, which was solution 1; 17 g (0.05 mol) of sucrose (sucrose was frozen in The final concentration in the preservation solution is 0.5mol L -1 ) ultrasonically dissolved in 25 mL of DPBS, after the sucrose was completely dissolved, add 10 mL of ethylene glycol and 10 mL of DMSO to obtain solution 2. After solution...
Embodiment 2
[0128] Example 2: Stem cell cryopreservation
[0129] Cryopreservation solution C1:
[0130] Each 100ml contains the following components:
[0131] substance Dosage PVA(g) 1.0 Ethylene glycol (mL) 10 Sucrose (mol L -1 )
0.5 Fetal bovine serum (mL) 20 DPBS(ml) margin
[0132] Cryopreservation Solution D:
[0133] Each 100ml contains the following components:
[0134] substance Dosage PVA(g) 2.0 Ethylene glycol (mL) 10 Sucrose (mol L -1 )
0.5 DPBS(mL) margin
[0135] Cryopreservation Solution F:
[0136] Each 100ml contains the following components:
[0137]
[0138]
[0139] The preparation method of cryopreservation solution is the same as that of Example 1.
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