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NKT cell preserving fluid and method

A technology of NKT cells and preservation methods, applied in the field of cell preservation, can solve the problems of killing activity affecting the therapeutic effect of NKT cells, high requirements for preservation equipment, and large damage to NKT cells, so as to achieve rapid recovery of activity, meet clinical needs, and prolong preservation the effect of time

Active Publication Date: 2022-01-21
广州希灵生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The survival rate and killing activity of NKT cells after preservation will affect the subsequent therapeutic effect of NKT cells
The common storage method is to store NKT cells at ultra-low temperature in liquid nitrogen. This method is suitable for long-term storage. There are many inconveniences in using liquid nitrogen to store cells that will be used only for a short period of time. First, liquid nitrogen storage requires storage equipment. Higher, inconvenient transportation, and the cooling process in liquid nitrogen storage greatly damages NKT cells, causing some cells to undergo apoptosis and affecting cell viability
The recovery of cell viability after storage in liquid nitrogen is slow

Method used

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  • NKT cell preserving fluid and method

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Experimental program
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Effect test

Embodiment 1

[0020] A kind of NKT cell preservation liquid, comprises solution medium and additive, and described solution medium is the mass concentration 0.9% sodium chloride solution, and described additive is: hydroxyethyl starch, D-mannitol, human serum albumin, cinnamamide, Meprobamate, zinc 2-hydroxysuccinate, L-glutamyl; the concentration of each component in the additive in the solution medium is: hydroxyethyl starch 12mg / mL, D-mannitol 23μg / mL, human serum white Protein 1mg / mL, cinnamamide 13μg / mL, meprobamate 0.4 mg / mL, zinc 2-hydroxysuccinate 1.5 mg / mL, L-glutamine 2mg / mL.

[0021] A method for preserving NKT cells, the process is as follows: NKT cells are resuspended with the above-mentioned preservation solution, and the cell density is 3×10 7 pcs / mL, divided into cryopreservation tubes, 1 mL per tube, and stored at 4°C.

Embodiment 2

[0023] A kind of NKT cell preservation liquid, comprises solution medium and additive, and described solution medium is the mass concentration 0.9% sodium chloride solution, and described additive is: hydroxyethyl starch, D-mannitol, human serum albumin, cinnamamide, Meprobamate, zinc 2-hydroxysuccinate, L-glutamyl; the concentration of each component in the additive in the solution medium is: hydroxyethyl starch 10mg / mL, D-mannitol 20μg / mL, human serum white Protein 0.5mg / mL, Cinnamamide 15μg / mL, Meprobamate 0.1 mg / mL, Zinc 2-Hydroxysuccinate 1mg / mL, L-Glutamine 1mg / mL.

[0024] A method for preserving NKT cells, the process is as follows: NKT cells are resuspended with the above-mentioned preservation solution, and the cell density is 1×10 7 pcs / mL, divided into cryopreservation tubes, 1 mL per tube, and stored at 6°C.

Embodiment 3

[0026] A kind of NKT cell preservation liquid, comprises solution medium and additive, and described solution medium is the mass concentration 0.9% sodium chloride solution, and described additive is: hydroxyethyl starch, D-mannitol, human serum albumin, cinnamamide, Meprobamate, zinc 2-hydroxysuccinate, L-glutamyl; the concentration of each component in the additive in the solution medium is: hydroxyethyl starch 15mg / mL, D-mannitol 25μg / mL, human serum white Protein 2mg / mL, Cinnamamide 20μg / mL, Meprobamate 0.5 mg / mL, Zinc 2-Hydroxysuccinate 2 mg / mL, L-Glutamine 4mg / mL.

[0027] A method for preserving NKT cells, the process is as follows: NKT cells are resuspended with the above-mentioned preservation solution, and the cell density is 4×10 7 pcs / mL, divided into cryopreservation tubes, 1 mL per tube, and stored at 4°C.

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Abstract

The invention discloses an NKT cell preserving fluid. The preserving fluid comprises a solution medium and additives, the solution medium is a sodium chloride solution with a mass concentration of 0.9%, and the additives are hydroxyethyl starch, D-mannitol, human serum albumin, cinnamamide, meprobamate, zinc 2-hydroxysuccinate and L-glutamine. According to the NKT cell preserving fluid provided by the invention, the components such as the cinnamamide, the meprobamate and the zinc 2-hydroxysuccinate added in the NKT cell preserving fluid have a synergistic effect, a stable preserving environment is provided for cells, temporary preservation of the NKT cells at 4-6 DEG C is realized, the preservation time of the NKT cells is prolonged, and the preserved NKT cells are high in viability, can quickly recover activity and fully meet clinical requirements. DMSO is not used in the preserving fluid, and the use amount of human serum albumin is reduced, so the safety of the preserving fluid is better. The invention further provides an NKT cells preserving method. The preserving fluid provided by the invention is adopted, the operation is simple and convenient, the preservation cost is effectively reduced, and convenience is provided for short-distance transportation of the NKT cells.

Description

technical field [0001] The invention relates to the field of cell preservation, in particular to a NKT cell preservation solution and a preservation method. Background technique [0002] Immune cells refer to cells that participate in the immune response or are related to the immune response. Various immune cells play an important role in the human body. NKT cells (natural killer T cells) are another new type of immune cells following T lymphocytes, B lymphocytes, and natural killer cells (NK cells). NKT cells are developed from the branch of thymic T cells, and are a group of immune cells with both T cell receptor TCR and NK cell characteristics on the cell surface. NKT cells have dual immunomodulatory effects of enhancing and suppressing immune responses. It connects the innate immune response and the acquired immune response, restricts recognition of CD1d-glycolipid antigens, and rapidly secretes a large number of cytokines after activation to regulate the body's immune...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 李颜军王国宇
Owner 广州希灵生物科技有限公司
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