A kind of cell preservation solution and its application in flow cytometry sample preservation

A technology of preservation solution and cells, which is applied in the field of preservation of cell samples, can solve problems such as poor preservation effect, misdiagnosis of leukemia/lymphoma immunophenotyping cases, decreased antigen expression intensity, etc., and achieve stable and correct results

Active Publication Date: 2017-09-12
ZHEJIANG BOZHEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, there are few studies on the samples for flow cytometry and their storage / processing methods in the prior art
U.S. Patent No. 5,753,428A discloses a cell preservation solution, but the inventors have found that, due to the use of citrate and other substances, the markers such as CD33 and CD13 on the blood sample are unstable, and the sample time preserved by it is slightly longer. Flow cytometric testing as described in Chinese Patent No. 201610027214 will produce different results from the original sample testing
There are few domestic studies on this. For example, Chinese Patent No. 201410534999 discloses a reagent for stabilizing blood sample cells. Although it claims to be used for flow cytometry detection, as an industry insider, it uses Potassium dichromate of strong zinc oxide, its preservation effect is not as good as the commonly used cell preservation solution containing paraformaldehyde
[0004] The inventor found that, especially the flow cytometry test described in the inventor's Chinese Patent No. 201610027214 involves the interpretation of the results by antibodies, so if the test cannot be performed in time, or the sample is processed in a suitable way, it will cause antigen expression A disproportionate decrease in intensity can greatly reduce the accuracy of the results and can even lead to misdiagnosis of leukemia / lymphoma immunophenotyping cases

Method used

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  • A kind of cell preservation solution and its application in flow cytometry sample preservation
  • A kind of cell preservation solution and its application in flow cytometry sample preservation
  • A kind of cell preservation solution and its application in flow cytometry sample preservation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Bottled cell preservation solution 1 and its preparation

[0040] Weigh 3.5814g Na2HPO4.12H2O, 0.272g KH2PO4, 8.0145g NaCl, 0.20115g KCl, 100g polyethylene glycol (PEG-20000), 5g chromium trichloride, dissolve in 900ml purified water, and adjust the volume to 1L. Filtered with a 7.25, 0.22 μm membrane, filled into a sterile reagent bottle, and sealed the reagent bottle to obtain cell preservation solution 1. After testing, it can be stored stably for at least 2 years at 2 to 8 °C, which is suitable for long-term store. For blood samples that have been added with anticoagulant, they can be used in aliquots.

Embodiment 2

[0041] Example 2 Bottled cell preservation solution 2 and its preparation

[0042] Weigh 3.5814g Na2HPO4.12H2O, 0.272gKH2PO4, 8.0145g NaCl, 0.20115g KCl, 100g polyethylene glycol (PEG-20000), 5g chromium trichloride, 15g EDTA.K2, dissolve in 900ml purified water, and set to volume To 1L, its pH is 7.25, filtered with 0.22μm membrane, filled into a sterile reagent bottle, and sealed the reagent bottle to obtain cell preservation solution 1. After testing, it can be stored stably at 2-8°C for at least 18 months, suitable for longer-term storage. For the original unanticoagulated blood sample, it can be used directly in aliquots.

Embodiment 3

[0043] Embodiment 3 Non-vacuum tube packed cell preservation solution 3 and its preparation

[0044] The cell preservation solution 1 prepared in Example 2 was taken, filled into sterile plastic tubes, the filling volume of each tube was 200 μl, and the tube stopper was sealed, thereby the cell preservation solution 3 was prepared. It can be used directly.

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PUM

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Abstract

The invention provides the application of a cell preservation liquid in flow cytometry detection. During detection, the typing of leukemia and lymphoma is completed. The cell preservation liquid comprises Na2HPO4, KH2PO4, NaCl, KCl, polyethylene glycol, chromium chloride and any optionally selected anticoagulant. In addition, the invention also discloses a method for preparing the cell preservation liquid and a method for preserving a flow-type cell sample. The cell preservation liquid can be used for the long-time and stable preservation of clinical cell samples. Especially for flow-type cell samples, the long-term and stable preservation effect is realized.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular, the invention relates to a preservation method of a cell-containing sample detected by a flow cytometer. Background technique [0002] With the development of biological and medical technology, scientific research detection and clinical diagnosis have gradually risen to the level of cells and genes. As one of the important means of quantitative analysis of single cells or biological ions, flow cytometry is also widely used. The working principle of flow cytometry is the multi-parameter and rapid quantitative analysis of single cells or other biological particles by monoclonal antibodies at the cellular and molecular level. It can analyze tens of thousands of cells at high speed, and can measure multiple parameters from one cell at the same time. It has the advantages of fast speed, high precision and good accuracy. It is one of the most advanced cell quantitative analysis tech...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/28G01N33/533G01N33/574
CPCA01N1/0226G01N1/28G01N33/533G01N33/57407G01N33/57426
Inventor 倪万茂
Owner ZHEJIANG BOZHEN BIOTECH CO LTD
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