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T lymphocyte cryopreservation solution

A lymphocyte and cryopreservation technology, applied in the field of cell biology, can solve the problems of high sulfone content, increased complexity, and increased risk of clinical use of frozen liquid, so as to achieve high survival rate, reduce risk, and maintain The effect of biological function

Active Publication Date: 2021-06-01
朱灏
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] CN111657264A provides a kind of T lymphocyte cryopreservation liquid and its preparation method and cell cryopreservation method, the freezing liquid composition includes dimethyl sulfoxide, serum-free cell culture medium, serum substitute, but the content of dimethyl sulfoxide High, due to the toxic effect of dimethyl sulfoxide, the freezing liquid with high content of dimethyl sulfoxide increases the risk of clinical use
In addition, when cryopreserved cells are used clinically, they need to be centrifuged to remove the components of the cryopreserved solution, which increases the complexity of use

Method used

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  • T lymphocyte cryopreservation solution

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preparation example Construction

[0035] A second aspect of the present invention provides a method for preparing a T lymphocyte cryopreservation solution, comprising the following steps:

[0036] S1. According to the formula, absorb DMSO and add it to a 500mL receiver, then slowly add compound electrolyte injection, dextran 40 glucose injection, and Duchenne's medium into it, and shake until mixed;

[0037] S2, then slowly add the formula amount of human serum albumin, and mix upside down to obtain the T lymphocyte cryopreservation solution.

[0038] The method also solves the phenomenon that human serum albumin encounters DMSO to cause protein precipitation.

[0039] In order to reduce the damage caused by DMSO in the freezing solution to the cells at room temperature, in some preferred embodiments, the preparation method of the T lymphocyte freezing solution also includes placing the configured T lymphocyte freezing solution in a medical refrigerator Pre-cool for more than 30 minutes.

[0040] As a prefer...

Embodiment 1

[0050] Embodiment 1 provides a kind of T lymphocyte cryopreservation solution, its preparation raw material includes 5 parts of DMSO, 56 parts of compound electrolyte injection, 8.5 parts of dextran 40 glucose injection, 25 parts of human albumin by volume. , 10 copies of Duchenne's modified medium.

[0051] The DMSO is a clinical grade DMSO purchased from WAK-Chemie;

[0052] The compound electrolyte injection was purchased from Hebei Tiancheng Pharmaceutical Co., Ltd.;

[0053] The dextran 40 glucose injection was purchased from Sichuan Kelun Pharmaceutical Co., Ltd.;

[0054] The human albumin was purchased from Octapharma AB;

[0055] The Duchenne's modified medium contains pyridoxine hydrochloride, purchased from Shanghai Hengfei Biotechnology Co., Ltd., model D5546.

[0056] The second aspect of embodiment 1 provides a kind of preparation method of T lymphocyte cryopreservation liquid, comprises the following steps:

[0057] S1. According to the formula, absorb DMSO ...

Embodiment 2

[0064] Embodiment 2 provides a kind of T lymphocyte cryopreservation liquid, its preparation raw material includes 5 parts of DMSO, 56 parts of compound electrolyte injection, 8.5 parts of dextran 40 glucose injection, 25 parts of human albumin by volume , 10 copies of Duchenne medium.

[0065] The DMSO is a clinical grade DMSO purchased from WAK-Chemie;

[0066] The compound electrolyte injection was purchased from Hebei Tiancheng Pharmaceutical Co., Ltd.;

[0067] The dextran 40 glucose injection was purchased from Sichuan Kelun Pharmaceutical Co., Ltd.;

[0068] The human albumin was purchased from Octapharma AB;

[0069] The Duchenne medium was purchased from Shanghai Hengfei Biotechnology Co., Ltd., model D5523.

[0070] The second aspect of embodiment 2 provides a kind of preparation method of T lymphocyte cryopreservation liquid, comprises the following steps:

[0071] S1. According to the formula, absorb DMSO and add it to a 500mL receiver, then slowly add compound...

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Abstract

The invention discloses a T lymphocyte cryopreservation solution, which is prepared from the following raw materials by volume: 1-5 parts of DMSO, 5-15 parts of dextran 40 glucose injection, 40-60 parts of compound electrolyte injection, 20-40 parts of human serum albumin and 5-15 parts of Dulbecco's medium or Dulbecco's modified medium. The invention also discloses a preparation method and a cryopreservation method thereof, the content of DMOS is reduced, so that the clinical use risk is reduced, the T lymphocyte still keeps a good cryopreservation effect, and the T lymphocyte has high survival rate and keeps a better form after cryopreservation and resuscitation. Besides, in the preparation process, the phenomenon that the human serum albumin generates protein precipitation when encountering DMSO is also solved, cells cryopreserved by the cryopreservation solution can be directly diluted and then applied clinically, components of the cryopreservation solution do not need to be centrifugally removed, and the cryopreservation solution can be used as an auxiliary material and is directly applied to clinical administration, so that the cryopreservation solution is more convenient to use.

Description

technical field [0001] The invention belongs to the technical field of cell biology, in particular to a T lymphocyte cryopreservation solution. Background technique [0002] The most commonly used technology for cell cryopreservation is liquid nitrogen cryopreservation, which mainly uses a slow freezing method with an appropriate amount of protective agent to freeze cells, such as glycerol or dimethyl sulfoxide (DMSO) as a protective agent, because if the cells are stored in Freezing directly without adding any protective agent, the water inside and outside the cells will quickly form ice crystals, which will cause a series of adverse reactions. [0003] T lymphocytes are derived from pluripotent stem cells of bone marrow. Mature T cells are distributed to the thymus-dependent areas of peripheral immune organs to settle in the peripheral immune organs through blood flow, and can be recirculated through lymphatic vessels, peripheral blood and tissue fluid to exert cellular im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0284
Inventor 朱灏
Owner 朱灏
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