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Balanced salt solution of marine invertebrate blood cells

A technology of invertebrates and balanced salt, which is applied to the preservation of invertebrate cells, animal cells, human or animal bodies, etc. It can solve the problems that cannot meet the needs of marine invertebrate cell preservation and subculture blood cell preservation, and achieve cell highly active effect

Active Publication Date: 2022-03-04
BEIBU GULF UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem that most of the current salt buffer solutions or balanced salt solutions including artificial seawater cannot meet the preservation and subculture of marine invertebrate cells, especially the preservation and subculture of blood cells, the present invention provides a marine invertebrate Balanced salt solution for blood cells, suitable for most marine invertebrate blood cells, with little damage to blood cells and high cell activity, which can protect marine invertebrate blood cells from lysing when stored in an isotonic solution

Method used

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  • Balanced salt solution of marine invertebrate blood cells
  • Balanced salt solution of marine invertebrate blood cells
  • Balanced salt solution of marine invertebrate blood cells

Examples

Experimental program
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Effect test

Embodiment 1

[0022] A balanced salt solution of marine invertebrate blood cells is prepared through the following steps:

[0023] 1) Dissolve 12mM KCl, 450mM NaCl, 20mM Na per liter of deionized water 2 HPO 4 , 10mM HEPES, penicillin 500U / ml, streptomycin 450μg / ul, nystatin 2μg / ml;

[0024] 2) adjust the pH of the solution obtained in the previous step to be 7.5, and filter and sterilize with a 0.22 μm filter membrane to obtain the balanced salt solution;

[0025] The marine invertebrates are selected from Hepu Pearl Oyster; step 1)-step 2) is to prepare the balanced salt solution now.

[0026] experiment procedure:

[0027] Use a 1mL syringe to draw 2mL of blood from the adductor muscle of Hepu pinnacle, and immediately add pre-cooled 3 times the volume of PBS buffer, artificial seawater (26.726g / L NaCl, 2.260g / L Mgcl 2 , 3.248g / L MgS0 4 , 1.153g / LCaCl 2 , NaHC0 3 0.198g / L, 0.743g / L KCl, 0.051g / L NaBr, 0.053g / L H 3 B0 3 , 0.0024g / LNa 2 SiO 3 , 0.0015g / L Na 2 Si 4 o 9 , 0.00...

Embodiment 2

[0033] A balanced salt solution of marine invertebrate blood cells is prepared through the following steps:

[0034] 1) Dissolve 10mM KCl, 500mM NaCl, 23mM Na per liter of deionized water 2 HPO 4 , 13mM HEPES, 0.05% (w / w) BSA, penicillin 550U / ml, streptomycin 600μg / ul, nystatin 4μg / ml;

[0035] 2) adjust the pH of the solution obtained in the previous step to be 7.0, and filter and sterilize with a 0.22 μm filter membrane to obtain the balanced salt solution;

[0036] The marine invertebrates are selected from abalone; step 1)-step 2) are used to prepare the balanced salt solution.

[0037] experiment procedure:

[0038] Extract 2mL of blood at the adductor muscle of abalone with a 1mL syringe, and the rest of the steps are the same as in Example 1;

[0039] Experimental results: As shown in Table 2, the survival rate of blood cells in the abalone blood cell balanced salt solution group was significantly higher than that in the PBS group and the artificial seawater group. ...

Embodiment 3

[0043] A balanced salt solution of marine invertebrate blood cells is prepared through the following steps:

[0044] 1) Dissolve 15mM KCl, 400mM NaCl, 20mM Na per liter of deionized water 2 HPO 4 , 12mM HEPES, 0.1% (w / w) BSA, penicillin 600U / ml, streptomycin 400μg / ul, nystatin 3μg / ml;

[0045] 2) adjust the pH of the solution obtained in the previous step to be 7.5, and filter and sterilize with a 0.22 μm filter membrane to obtain the balanced salt solution;

[0046] The marine invertebrates are selected from clams; step 1)-step 2) are used to prepare the balanced salt solution.

[0047] experiment procedure:

[0048] Take 2 mL of blood at the clam adductor muscle with a 1 mL syringe, and the rest of the steps are the same as in Example 1;

[0049] Experimental results:

[0050] As shown in Table 3, the survival rate of blood cells in clam blood cells in balanced salt solution group was significantly higher than that in PBS group and artificial seawater group.

[0051] T...

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Abstract

The invention discloses a balanced salt solution of marine invertebrate blood cells. The balanced salt solution is prepared by the following steps: 1) dissolving 10-15 mM of KCl, 400-500 mM of NaCl, 20-23 mM of Na2HPO4, 10-13 mM of HEPES, 500-700 U / ml of penicillin, 400-600 [mu] g / [mu] l of streptomycin and 2-4 [mu] g / ml of nystatin in each liter of deionized water; and 2) adjusting the pH value of the solution obtained in the step 1) to 7.0-7.5, and filtering and sterilizing with a 0.22 [mu] m filter membrane to obtain the balanced salt solution. The method is suitable for most marine invertebrate blood cells, the damage degree to the blood cells is small, the cell activity is high, and the marine invertebrate blood cells can be protected from cracking when stored in an isotonic solution.

Description

【Technical field】 [0001] The invention relates to the technical field of marine invertebrate cell biology, in particular to a balanced salt solution of marine invertebrate blood cells. 【Background technique】 [0002] A balanced salt solution is a salt solution that contains electrolytes similar to those in blood plasma. Balanced salt solution is mainly composed of inorganic salts and glucose. Its function is to maintain cell osmotic pressure balance, maintain pH stability and provide simple nutrition. The simplest balanced salt solution (BSS, balanced salt solution) is Ringer, which is to add potassium chloride and calcium chloride to normal saline, also known as Ringer's solution. Because it was invented by the British physiologist "Ringer", it is called Ringer's solution, which is actually the commonly known compound sodium chloride injection. Ringer's Solution is also called Compound Sodium Chloride Injection, because Ringer's Solution contains sodium ions, potassium io...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07A01N1/02
CPCC12N5/0601A01N1/0221A01N1/0215C12N2500/12C12N2500/60C12N2501/998
Inventor 喻达辉王培白丽蓉李素萍郑玉斯何积翠郭颖
Owner BEIBU GULF UNIV
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