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Immune cell cryopreservation liquid and cryopreservation method

A technology of immune cells and cryopreservation solution, which is applied in the field of cell biology, can solve the problems that the toxicity of cell cryopreservation solution easily causes allergic reactions and cannot be reinfused clinically, and achieves improved cryopreservation effect, high clinical safety, and simple formula Effect

Active Publication Date: 2018-06-12
重庆斯德姆生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the above-mentioned deficiencies in the prior art, the present invention provides an immune cell cryopreservation solution and a cryopreservation method, which can effectively solve the problem that the cell cryopreservation solution in the prior art has certain toxicity and is easy to cause allergic reactions, and cannot be directly used in clinical practice. Return problem

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] An immune cell cryopreservation solution, including a basal medium and supplementary components, wherein the basal medium is a DMEM / F12 medium, and the supplementary components include the following final concentrations of components: trehalose 1g / mL, propylene glycol 3v / v%, Acetamide 3v / v%, dextran 5v / v%, hydroxyethyl starch 1g / mL, glucose 0.5g / mL, heparin sodium 50U / mL, pearl grass extract 5mg / mL and BCG complex polysaccharide 0.5mg / mL .

[0024] Among them, the pearl grass extract is obtained as follows: take the pearl grass, clean it, dry it, grind it to 100 mesh, mix the powder with absolute ethanol at a weight ratio of 1:1, and then carry out CO 2 Supercritical extraction, finally centrifuged to remove impurities, to obtain a fat-soluble extract, add 6 times the weight of distilled water to the extraction residue, heat at 90°C for 8 hours, then filter, concentrate, and finally vacuum freeze-dry to obtain a water-soluble extract, combine the two The second extract...

Embodiment 2

[0027] An immune cell cryopreservation solution, including a basal medium and supplementary components, wherein the basal medium is a DMEM / F12 medium, and the supplementary components include components with the following final concentrations: trehalose 5g / mL, propylene glycol 6v / v%, Acetamide 5v / v%, dextran 10v / v%, hydroxyethyl starch 3g / mL, glucose 1.5g / mL, heparin sodium 150U / mL, pearl grass extract 15mg / mL and BCG complex polysaccharide 1.5mg / mL .

[0028] Among them, the pearl grass extract is obtained in this way: take the pearl grass, clean it, dry it, grind it to 100 mesh, mix the powder with absolute ethanol at a weight ratio of 3:1, and then carry out CO 2 Supercritical extraction, finally centrifuged to remove impurities, to obtain a fat-soluble extract, add 8 times the weight of distilled water to the extraction residue, heat at 95°C for 8h, then filter, concentrate, and finally vacuum freeze-dry to obtain a water-soluble extract, combine the two The second extrac...

Embodiment 3

[0031] An immune cell cryopreservation solution, including a basal medium and supplementary components, wherein the basal medium is a DMEM / F12 medium, and the supplementary components include components with the following final concentrations: trehalose 3g / mL, propylene glycol 3-6v / v %, acetamide 4v / v%, dextran 8v / v%, hydroxyethyl starch 2g / mL, glucose 1.0g / mL, heparin sodium 100U / mL, pearl grass extract 10mg / mL and BCG complex polysaccharide 0.8mg / mL.

[0032] Among them, the pearl grass extract is obtained in this way: take the pearl grass, wash it, dry it, grind it to 100 mesh, mix the powder with absolute ethanol at a weight ratio of 2:1, and then carry out CO 2 Supercritical extraction, finally centrifuged to remove impurities, to obtain a fat-soluble extract, add 8 times the weight of distilled water to the extraction residue, heat at 90°C for 8 hours, then filter, concentrate, and finally vacuum freeze-dry to obtain a water-soluble extract, combine the two The second ...

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PUM

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Abstract

The invention discloses an immune cell cryopreservation liquid and a cryopreservation method. The cryopreservation liquid comprises a basic culture medium and additives. The additives include, by final concentration, 1-5 g / mL of trehalose, 3-6 v / v% of propylene glycol, 3-5 v / v% of acetamide, 5-10 v / v% of dextran, 1-3 g / mL of hydroxyethyl starch, 0.5-1.5 g / mL of glucose, 50-150 U / mL of heparin sodium, 5-15 mg / mL of herba phyllanthi urinariae extract and 0.5-1.5 mg / mL of bacillus Calmette-Guerin composite polysaccharides. The cryopreservation liquid is simple in formulation, interaction and synergistic effects of the components are achieved, endothelial progenitor cell cryopreservation effects can be evidently improved by the adoption of the cryopreservation method, insusceptibility of post-reviving cell proliferation activity can be guaranteed, and it is guaranteed that intracellular moisture of cells close to a freezing point is free of crystallization.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to an immune cell cryopreservation solution and a cryopreservation method. Background technique [0002] Immune cell therapy is a new type of autoimmune anti-cancer treatment. It uses biotechnology and biological agents to culture and amplify immune cells collected from patients in vitro and then reinfuse them back into patients to stimulate and enhance immunity. The body's own immune function, so as to achieve the purpose of treating tumors. [0003] With the rapid development of immune cell biology and immune molecular biology, somatic cell immunotherapy has become one of the important means of adjuvant therapy after radiotherapy and chemotherapy for tumor patients. All have good effects. [0004] However, the effect of in vitro culture of immune cells depends largely on the number and quality of immune cells in the patient, but the number of immune cells in tum...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 王健熊华强秦连荣熊荣骞
Owner 重庆斯德姆生物技术有限公司
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