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Preparation method of interstitial fluid

A technology of tissue fluid and blood, which is applied in the field of biomedical experiments, can solve the problems of limited amount of extracted tissue fluid, large difference between the composition of tissue simulation fluid and real tissue fluid, etc., and achieve low production costs, meet the needs of biomedical experiments, and achieve the effect of mild conditions

Inactive Publication Date: 2020-05-19
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem that the composition of the existing tissue simulation fluid is very different from the real tissue fluid, and the amount of tissue fluid extracted from the body tissue is limited, and provides a preparation method that is closer to the tissue fluid, and the obtained tissue fluid is closer to the real tissue fluid

Method used

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  • Preparation method of interstitial fluid

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preparation example Construction

[0017] In order to achieve the purpose of the present invention, an embodiment of the present invention provides a method for preparing interstitial fluid, comprising the following steps:

[0018] Step 1. Collect blood from healthy people or experimental animals;

[0019] Step 2, subjecting the blood to anticoagulant or non-anticoagulant treatment, and centrifuging to obtain plasma or serum;

[0020] Step 3, add the serum or plasma to the filter of the ultrafiltration tube, insert the filter into the nested tube, perform centrifugation, collect the liquid in the nested tube, and obtain the pretreatment solution;

[0021] Step 4, using sodium citrate buffer solution to adjust the pH value of the pretreatment solution to 7.1-7.3 to obtain preliminary interstitial fluid;

[0022] Step 5: Filter and sterilize the preliminary interstitial fluid with a 0.22 micron microporous membrane to obtain the interstitial fluid.

[0023] Specifically, the process of collecting blood from hea...

Embodiment 1

[0031] 1. Collect blood from healthy people

[0032] 4 mL of fresh blood from healthy people was collected by blood anticoagulant tubes with EDTA anticoagulant, and the blood collection process was kept aseptic.

[0033] 2. Separation of plasma

[0034] The collected anticoagulant blood was centrifuged at a speed of 3000 rpm for 5 minutes. After the centrifugation was completed, the supernatant was collected with a pipette gun to obtain plasma.

[0035] 3. Separation of plasma proteins

[0036] Plasma was added to a 30kD molecular weight ultrafiltration tube, and the ultrafiltration tube was nested into the nested tube for high-speed centrifugation at 12,000 rpm for 10 minutes. After centrifugation, the liquid in the nested tube was collected to obtain preliminary interstitial fluid.

[0037] 4. Adjust the pH value

[0038] Use a pH meter to detect the pH value of the initially obtained interstitial fluid, add an appropriate amount of citrate buffer to it, and adjust the p...

Embodiment 2

[0042] 1. Collect blood from healthy people

[0043] 4mL of fresh blood from healthy people was collected using common blood collection tubes, and the blood collection process was kept aseptic.

[0044] 2. Separation of serum

[0045] After the collected blood was allowed to stand at room temperature for 3 hours, it was centrifuged at a speed of 3000 rpm for 5 minutes. After the centrifugation was completed, the supernatant was collected with a pipette gun to obtain serum.

[0046] 3. Separation of plasma proteins

[0047] Serum was added to a 30kD molecular weight ultrafiltration tube, the ultrafiltration tube was nested into the nested tube, and high-speed centrifugation was performed at a speed of 12,000 rpm for 10 minutes. After centrifugation, the liquid in the nested tube was collected to obtain preliminary interstitial fluid.

[0048] 4. Adjust the pH value

[0049] Use a pH meter to detect the pH value of the initially obtained interstitial fluid, add an appropriat...

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Abstract

The invention is suitable for the field of biomedical experiments, and provides a preparation method of an interstitial fluid. The method comprises the following steps: step 1, collecting blood of a healthy person or an experimental animal; step 2, carrying out anticoagulation or non-anticoagulation treatment on the blood, and performing centrifuging to obtain plasma or serum; step 3, adding the serum or the plasma into a filter of an ultrafiltration tube, embedding the filter into a nested tube, carrying out centrifugal operation, and collecting a liquid in the nested tube to obtain a pretreated liquid; step 4, adjusting the pH value of the pretreated liquid to 7.1-7.3 by using a sodium citrate buffer solution to obtain a primary interstitial fluid; and step 5, filtering and sterilizing the preliminary interstitial fluid by using a 0.22-micron microporous filter membrane to obtain the interstitial fluid. Components of the interstitial fluid obtained through the method are close to those of real interstitial fluid, and the interstitial fluid can meet biomedical experiment requirement and has good safety.

Description

technical field [0001] The invention belongs to the field of biomedical experiments, in particular to a method for preparing interstitial fluid. Background technique [0002] Interstitial fluid is the fluid that exists between cells, also known as interstitial fluid. Interstitial fluid exists in large quantities in animals and plants, and is the liquid environment in which ordinary cells live directly, and can exchange substances with cells. Cells infiltrate in interstitial fluid and can exchange substances with interstitial fluid, which is a necessary condition for normal cell metabolism. In addition, interstitial fluid can also buffer and protect cells. [0003] In animals, interstitial fluid infiltrates from the arterial end of the capillary into a part of the interstitial fluid, exchanges substances with the tissue cells, and then flows back into the blood or lymph through the venous end of the capillary or lymphatic capillary. Most of the interstitial fluid is in a g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078
CPCC12N5/0634
Inventor 唐敬龙郑玉新李燕婷
Owner QINGDAO UNIV
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