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Protein-free liquid-nitrogen cell frozen preservation liquid and preparation method thereof

A protein-free, cryopreservation solution technology, applied in the biological field, can solve problems such as difficulty in the source of human albumin, contamination of human-derived pathogens, and contamination of animal pathogens, and achieve the effects of high cell recovery rate, simplified operation, and low production cost

Inactive Publication Date: 2015-12-23
南京三生生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Cell cryopreservation technology is a very important means in cell biology technology. In the conventional cell cryopreservation solution, it is often necessary to add a certain amount of animal serum or at least human albumin to achieve a better cryopreservation effect. Serum may be contaminated by animal pathogens, the source of human albumin is difficult and costly, and there is also the possibility of contamination of human pathogens

Method used

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  • Protein-free liquid-nitrogen cell frozen preservation liquid and preparation method thereof

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Effect test

Embodiment 1

[0020] 1. Preparation of protein-free liquid nitrogen cell cryopreservation medium

[0021] A preparation method for protein-free liquid nitrogen cell freezing liquid, comprising the following steps:

[0022] (1) Weigh 6.0 g of dextran and dissolve it in 70 ml of RPMI-1640 cell basal medium;

[0023] (2) Add 1.0 g of poloxamer 188 to the above solution;

[0024] (3) Add 10ml DMSO, then add RPMI-1640 cell basal medium to make the volume to 100ml;

[0025] 2. The preparation and preparation method of the protein-free liquid nitrogen cell cryopreservation solution, the above-mentioned mixed solution is first clarified and filtered with a 0.45 μm filter membrane under non-sterile conditions, and then used a 0.22 μm filter membrane under sterile conditions. Sterilize by filtration to obtain a protein-free liquid nitrogen cell cryopreservation solution.

Embodiment 2

[0027] 1. Preparation of protein-free liquid nitrogen cell cryopreservation medium

[0028] A preparation method for protein-free liquid nitrogen cell freezing liquid, comprising the following steps:

[0029] (1) Weigh 4.0 g of dextran and dissolve it in 70 ml of RPMI-1640 cell basal medium;

[0030] (2) Add 2.0 g of poloxamer 188 to the above solution;

[0031] (3) Add 10ml DMSO, then add RPMI-1640 cell basal medium to make the volume to 100ml;

[0032] 2. The preparation and preparation method of the protein-free liquid nitrogen cell cryopreservation solution, the above-mentioned mixed solution is first clarified and filtered with a 0.45 μm filter membrane under non-sterile conditions, and then used a 0.22 μm filter membrane under sterile conditions. Sterilize by filtration to obtain a protein-free liquid nitrogen cell cryopreservation solution.

Embodiment 3

[0034] 1. Preparation of protein-free liquid nitrogen cell cryopreservation medium

[0035] A preparation method for protein-free liquid nitrogen cell freezing liquid, comprising the following steps:

[0036] (1) Weigh 12.0 g of dextran and dissolve it in 70 ml of RPMI-1640 cell basal medium;

[0037] (2) Add 0.5 g of poloxamer 188 to the above solution;

[0038] (3) Add 10ml DMSO, then add RPMI-1640 cell basal medium to make the volume to 100ml;

[0039] 2. The preparation and preparation method of the protein-free liquid nitrogen cell cryopreservation solution, the above-mentioned mixed solution is first clarified and filtered with a 0.45 μm filter membrane under non-sterile conditions, and then used a 0.22 μm filter membrane under sterile conditions. Sterilize by filtration to obtain a protein-free liquid nitrogen cell cryopreservation solution.

[0040] Contrast cell freezing solution (containing protein)

[0041] 1. Formula and preparation method of cryopreservation s...

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Abstract

The invention belongs to the technical field of biology and particularly relates to protein-free liquid-nitrogen cell frozen preservation liquid and a preparation method thereof. Each 100ml of protein-free liquid-nitrogen cell frozen preservation liquid contains 2.0-15.0g of any one of dextran, saccharosan and hydroxyethyl starch, 0.2-2.0g of poloxamer 188, 10ml of DMSO (Dimethylsulfoxide) and the balance of an RPMI-1640 (Roswell Park Memorial Institute-1640) cell basic culture medium. The protein-free liquid-nitrogen cell frozen preservation liquid provided by the invention is prepared from chemical components and does not contain protein; and the recovery rate of cells is high and the stability is good.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a protein-free liquid nitrogen cell cryopreservation solution and a preparation method and application method thereof. Background technique [0002] Cell cryopreservation technology is a very important means in cell biology technology. In the conventional cell cryopreservation solution, it is often necessary to add a certain amount of animal serum or at least human albumin to achieve a better cryopreservation effect. Serum may be contaminated by animal pathogens, the source of human albumin is difficult and costly, and there is also the possibility of contamination by human pathogens. At present, the use of cell cryopreservation technology has gone far beyond the scope of scientific research, especially in recent years, with the clinical promotion of cell therapy technology and stem cell technology, there is an urgent need for safer cell cryopreservation solutions in clin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 徐念沁
Owner 南京三生生物技术股份有限公司
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