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Animal lymphocyte separation liquid and preparation method thereof

A technology of lymphocyte and separation liquid, which is applied in the field of animal lymphocyte separation liquid and its preparation, can solve the problems of prolongation of cell band formation time, and achieve the effects of no platelet contamination, significantly reduced time, and good quality of lymphocytes

Active Publication Date: 2016-04-27
南京三生生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But at the same time, we also found that after substituting diatrizoate meglumine with sodium diatrizoate, the formation time of cell bands during centrifugation (cell banding) was significantly prolonged during lymphocyte separation.

Method used

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  • Animal lymphocyte separation liquid and preparation method thereof
  • Animal lymphocyte separation liquid and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0017] The traditional lymphocyte separation solution contains 10% diatrizoate and 6% polysucrose. Replacing expensive diatrizoate meglumine with lower-priced sodium diazoate, it is formulated as a low-cost lymphocyte separation solution containing 10% sodium diazoate and 6% polysucrose, and the separated lymphocytes The total number of cells is similar to that of the traditional formula of diatrizoate, but the time for the cells to form bands (banding time) during the cell separation process is significantly prolonged, as shown in Table 1.

Embodiment 2

[0019] The present invention improves the formula, keeps the concentration of 10% sodium diatrizoate unchanged, replaces part of polysucrose with polyvinylpyrrolidone (PVP) to configure a new lymphocyte separation fluid, and the total concentration of polysucrose and polyvinylpyrrolidone is 6 %, for lymphocyte separation.

[0020] The further improved formula of the separating liquid is 10% sodium diatrizoate, 5.2% polysucrose, 0.8% polyvinylpyrrolidone, and the balance is pyrogen-free deionized water, the total amount is 100%, and the separating liquid 1 is obtained. For the separation of lymphocytes, the separation effect is shown in Table 2.

Embodiment 3

[0022] The further improved formula of the separating liquid is 10% sodium diatrizoate, 5% polysucrose, 1.0% polyvinylpyrrolidone, and the balance is pyrogen-free deionized water, the total amount is 100%, and the separating liquid 2 is obtained. For the separation of lymphocytes, the separation effect is shown in Table 2.

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Abstract

The invention discloses animal lymphocyte separation liquid, which consists of the following components in percentage by mass: 10% of sodium diatrizoate, 5.2-4.4% of ficoll, 0.8-1.6% of a hydrophilic high-molecular polymer of non-glycosyl units and the balance of de-ionized water free from a heat source, totaling 100%, wherein the total concentration of the ficoll and the hydrophilic high-molecular polymer of the non-glycosyl units is 6%; and the specific weight of the animal lymphocyte separation liquid is 1.076-1.078g / ml at 20 DEG C. The animal lymphocyte separation liquid, which is low in cost and high in efficiency, is prepared by replacing expensive meglumine diatrizoate with sodium diatrizoate and by mixing and combining the sodium diatrizoate with the ficoll, and on the basis of a consistent separation effect of the sodium diatrizoate with the meglumine diatrizoate, the production cost is significantly reduced; and by adding the hydrophilic high-molecular polymer of the non-glycosyl units, such as polyvinylpyrrolidone, polyvinyl alcohol and polypropylene glycol, the cell banding time of separating lymphocytes with the low-cost prescription is significantly shortened, meanwhile the quality of the obtained lymphocytes is significantly improved, platelet contamination is nearly avoided.

Description

technical field [0001] The invention relates to an animal lymphocyte separation liquid and a preparation method thereof, belonging to the technical field of biomedicine. Background technique [0002] The traditional animal lymphocyte separation solution is configured with diatrizoate and polysucrose. The principle is to rely on polysucrose to form a concentration gradient under centrifugation conditions to achieve the separation of different cells, but because the viscosity of polysucrose is very high, it is not conducive to cell movement during the separation process. In order to improve the viscosity of polysucrose, often in A certain amount of diatrizoate meglumine is added to the formula to reduce the viscosity and facilitate the movement of cells during the separation process. But the price of diatrizoate meglumine is very expensive. We try to replace diatrizoate meglumine with sodium diatrizoate, and the price will be reduced by 6 times. Our preliminary test results ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781C12N5/0783
CPCC12N5/0635C12N5/0636C12N2500/30C12N2500/34C12N2500/50
Inventor 郁庆明靖志安田莉姜强徐炜华徐念沁
Owner 南京三生生物技术股份有限公司
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