Freezing preservation solution used for adipose-derived stem cells as well as preparation method for freezing preservation solution and culture medium

A culture medium and amino acid technology, applied in the field of cryopreservation solution, can solve the problems of lack of serum-free medium research, etc., and achieve the effect of enhancing cryopreservation effect, promoting protein expression, enhancing cryopreservation effect and preservation effect

Inactive Publication Date: 2017-04-19
浙江译美生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Based on the in-depth study of this serum-free cryopreservation solution, the inventor realized that although this serum-free cryopreservation solution has a good cryopreservation effect, this serum-free cryopreservation solution only provides a specific serum-free cryopreservation solution formulated with KSR. Serum-free medium in serum-free cryopreservation was not further studied

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  • Freezing preservation solution used for adipose-derived stem cells as well as preparation method for freezing preservation solution and culture medium
  • Freezing preservation solution used for adipose-derived stem cells as well as preparation method for freezing preservation solution and culture medium

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Effect test

Embodiment 1

[0038] A method for preparing a cryopreservation solution for adipose-derived stem cells, comprising the following preparation steps:

[0039] Step1: According to parts by weight, weigh 7200 parts of inorganic salt complex, 990 parts of amino acid compound, 40 parts of vitamin compound, 1000 parts of glucose, 110 parts of sodium pyruvate, 68 parts of butyric acid, 80 parts of sodium butyrate, citric acid 0.15 parts of iron, 1,000,000 parts of water, 9 parts of phenol red, 1 part of fibronectin, 25 parts of hormone complex, and 50,000 parts of penicillin, mixed and stirred evenly to obtain a culture medium;

[0040] Step2: Take 8% dimethyl sulfoxide, 77% serum replacement component KSR and 15% medium according to the volume fraction, mix and stir evenly.

[0041] Wherein the hormone complex includes insulin, glucagon and progesterone, and according to the weight ratio, the insulin:glucagon:progesterone=3:3:1.

Embodiment 2

[0043] A method for preparing a cryopreservation solution for adipose-derived stem cells, comprising the following preparation steps:

[0044] Step1: In parts by weight, weigh 7230 parts of inorganic salt complex, 980 parts of amino acid compound, 41 parts of vitamin compound, 1030 parts of glucose, 107 parts of sodium pyruvate, 60 parts of butyric acid, 85 parts of sodium butyrate, citric acid 0.22 parts of iron, 1,000,000 parts of water, 9.2 parts of phenol red, 0.8 parts of fibronectin, 20 parts of hormone complex, and 50,000 parts of streptomycin were mixed and stirred evenly to obtain a culture medium;

[0045] Step2: Take 14% dimethyl sulfoxide, 74% serum replacement component KSR and 12% medium according to the volume fraction, mix and stir evenly.

[0046] Wherein the hormone complex includes insulin, glucagon and progesterone, and according to the weight ratio, the insulin:glucagon:progesterone=3:5:1.

Embodiment 3

[0048] A method for preparing a cryopreservation solution for adipose-derived stem cells, comprising the following preparation steps:

[0049] Step1: In parts by weight, weigh 7260 parts of inorganic salt complex, 985 parts of amino acid compound, 43 parts of vitamin compound, 1060 parts of glucose, 105 parts of sodium pyruvate, 65 parts of butyric acid, 90 parts of sodium butyrate, citric acid 0.18 parts of iron, 1,000,000 parts of water, 9.4 parts of phenol red, 0.6 parts of fibronectin, 15 parts of hormone complex, and 50,000 parts of penicillin were mixed and stirred evenly to obtain a culture medium;

[0050] Step2: Take 20% dimethyl sulfoxide, 70% serum replacement component KSR and 10% medium according to the volume fraction, mix and stir evenly.

[0051] Wherein the hormone complex includes insulin, glucagon and progesterone, and according to the weight ratio, the insulin:glucagon:progesterone=4:3:1.

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Abstract

The invention discloses a freezing preservation solution used for adipose-derived stem cells as well as a preparation method for the freezing preservation solution and a culture medium. The culture medium comprises the following components in parts by weight: 7200-7300 parts of inorganic salt composite, 980-1000 parts of amino acid complex, 40-45 parts of vitamin composite, 1000-1100 parts of glucose, 100-110 parts of sodium pyruvate, 60-75 parts of butyric acid, 80-110 parts of sodium butyrate, 0.15-0.22 part of ferric citrate and 1,000,000 parts of water. The freezing preservation solution has the beneficial effects that: the culture medium has excellent freezing preservation effect in the freezing preservation solution which is compounded by a serum substitute component KSR and dimethyl sulfoxide, and is used for the adipose-derived stem cells.

Description

technical field [0001] The invention relates to the field of cryopreservation liquid, in particular to a cryopreservation liquid for adipose stem cells, a preparation method thereof and a culture medium. Background technique [0002] Bone marrow stem cells, umbilical cord stem cells and adipose-derived stem cells (ADSCs) are several types of stem cells that have been studied more in recent years. Bone marrow stem cells are difficult to obtain, slow to expand, unsuitable for frail and elderly patients, and ethical issues, which bring inconvenience to clinical application. Compared with bone marrow stem cells, umbilical cord mesenchymal stem cells have a wide range of sources, low immunogenicity, and almost no immune rejection. Adipose stem cells are a kind of stem cells with multi-directional differentiation potential isolated from adipose tissue in recent years. Compared with bone marrow stem cells and umbilical cord stem cells, ADSCs have many advantages. Studies have foun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226
Inventor 陈继冰吴振化
Owner 浙江译美生物科技有限公司
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