Cell freezing medium, freezing resuscitation method and application thereof
A cryopreservation, cell suspension technology, applied in the field of clinical medicine, can solve problems such as cell damage, and achieve the effects of reducing damage, reducing cell damage, and improving cell activity
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Embodiment 1
[0107] The cell cryopreservation solution of this embodiment and its preparation method include: the preparation of the equilibrium buffer, the preparation of the acylated ε-polylysine solution, and the preparation of the equilibrium buffer and the acylated ε-polylysine solution The preparation of the main cell cryopreservation solution will be described in detail later.
[0108] 1. The preparation of the equilibrium buffer includes the following steps: add NaCl 100mmol, KCl 10mmol, MgCl to ultrapure water in sequence 2 13mmol, CaCl 2 0.25mmol, after all the above substances are completely dissolved, adjust the pH of the above solution to 7.0~7.6 with 10mol / L NaOH aqueous solution; then dilute to 1L with ultrapure water.
[0109] 2. Preparation steps of acylated ε-polylysine solution: Weigh 13g of succinic anhydride and pour it into 100mL of ε-polylysine aqueous solution with a mass concentration of 0.25g / mL, and put it into 50°C water for water bath , after 2 hours of water...
Embodiment 2
[0116] The cell cryopreservation solution of this embodiment and its preparation method include: the preparation of the equilibrium buffer, the preparation of the acylated ε-polylysine solution, and the preparation of the equilibrium buffer and the acylated ε-polylysine solution The preparation of the main cell cryopreservation solution will be described in detail later.
[0117] 1. The preparation of the equilibrating buffer comprises the following steps:
[0118] Step A: Add NaNO sequentially to ultrapure water 3 15mmol, KNO 3 10mmol, Mg(NO 3 ) 2 10mmol, Ca(NO 3 ) 2 0.5mmol, dissolve all the above substances completely;
[0119] Step B: Add 15mmol of mannitol, 1mmol of vitamin C, 3mmol of glutathione, 2mmol of N-acetyl-L-cysteine, 2mmol of tert-butylterephthalene, and 2 μmol of phenol, 20 mmol of histidine, 14 mmol of glutamic acid, 1 mmol of α-ketoglutarate, and 5 mmol of adenosine, until all substances are completely dissolved;
[0120] Step C: then adjust the pH o...
Embodiment 3
[0129] The cell cryopreservation solution of this embodiment and its preparation method include: the preparation of the equilibrium buffer, the preparation of the acylated ε-polylysine solution, and the preparation of the equilibrium buffer and the acylated ε-polylysine solution The preparation of the main cell cryopreservation solution will be described in detail later.
[0130] 1. The preparation of the equilibrating buffer comprises the following steps:
[0131] Step A1: Add the NaCl 100mmol, the KCl 10mmol, the MgCl to the ultrapure water successively 2 13mmol, the CaCl 2 0.25mmol, dissolve all the above substances completely;
[0132] Step B1: Add 10 mmol of mannitol, 0.5 mmol of vitamin C, 2 mmol of glutathione, 5 mmol of N-acetyl-L-cysteine, and 5 mmol of tert-butyl-p-phenylene in sequence. 5 μmol of diphenol, 24.5 mmol of histidine, 16.3 mmol of glutamic acid, 0.5 mmol of α-ketoglutaric acid, 3 mmol of adenosine and 15 g of human serum albumin, until all substances ...
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