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Circulating tumor cell separation and enrichment kit

A technology for separation and enrichment of tumor cells, applied in the field of circulating tumor cell separation and enrichment kits, can solve the problem of inability to guarantee the repeatability and reliability of CTCs analysis and detection, the inability to effectively protect the morphology and antigenic epitopes of CTCs, and the inconsistency of whole blood samples. Stability and other issues, to achieve good CTCs preservation effect, improve separation and enrichment efficiency and purity, and good red blood cell lysis effect

Active Publication Date: 2020-12-22
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing cell preservation solution still has the following problems: low temperature storage is required, but low temperature storage puts forward higher requirements on the stability and transportation conditions of the sample; it cannot be stored for a long time at room temperature, making the collected CTCs cells It is easily broken and degraded within hours, cannot effectively protect the morphology and antigenic epitopes of CTCs, and cannot guarantee the repeatability and reliability of CTCs analysis and detection, which is not conducive to subsequent analysis and detection
And the number of CTCs in the peripheral blood of patients is very small, which makes whole blood samples containing CTCs cells very unstable during transportation

Method used

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  • Circulating tumor cell separation and enrichment kit
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  • Circulating tumor cell separation and enrichment kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 A circulating tumor cell isolation and enrichment kit

[0045] The composition of a circulating tumor cell separation and enrichment kit in this embodiment is as follows:

[0046] 1. Circulating tumor cell separation and enrichment preservation solution:

[0047] The circulating tumor cell separation and enrichment preservation solution is an aqueous solution containing the following concentration components: NaCl3.5mmol / L, NH 4 Cl 13g / L, Tris-HCl 12.5mmol / L, 4-hydroxyethylpiperazineethanesulfonic acid 30mmol / L, corn oligopeptide 0.04w / v%, thimerosal 0.01w / v%.

[0048] The preparation method of the corn oligopeptide is as follows:

[0049] (1) Corn gluten solution configuration: Weigh commercially available corn gluten powder and place it in a reaction cup, add water to mix evenly according to the mass fraction of 10%, adjust the pH of the solution to 11 with sodium hydroxide, and the ultrasonic frequency is 40kHz, and the ultrasonic power is 45W , sonicate...

Embodiment 2

[0067] The preservation effect of embodiment 2CTCs

[0068] This example analyzes the preservation effect of the circulating tumor cell isolation and enrichment preservation solution in the kit described in Example 1 on the preservation of CTCs in biological samples. Use the cell preservation solutions of 4 different experimental groups in Table 1 below to store biological samples containing epithelial CTCs No. 1-3 at room temperature for 2 hours, and then use the detection described in the inventor’s previously published patent document (patent number: 2014102285119) The kit and detection steps detect the CTCs in the samples, and at the same time, the detection results of the biological samples that are directly detected without being stored in the cell preservation solution after sampling are used as the control.

[0069] Table 1 Cell Preservation Solution

[0070]

[0071] The test results of each group were as figure 1As shown, after comparison, it can be seen that af...

Embodiment 3

[0072] Embodiment 3 erythrocyte lysis efficiency

[0073] In order to analyze the lysing efficiency of circulating tumor cell separation and enrichment preservation solution on red blood cells in the sample in the present invention, 6 groups of blood samples were collected, and after initial counting of RBCs by an automatic blood cell counter, each group of blood samples was divided into 8 parts on average. The solutions of the 8 experimental groups in the following Table 2 were respectively taken for the erythrocyte lysis step (refer to step 1 in Example 1), and then the RBC was counted again by an automatic blood cell counter.

[0074] Table 2 each experimental group solution formula

[0075]

[0076] Each group of solutions in the above table weighed the components according to the formula and dissolved them in 1000mL of distilled water, adjusted the pH to 7.2-7.4, sterilized with a 0.22μm filter membrane, and labeled for use.

[0077] The detection results of the eryth...

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Abstract

The present invention provides a circulating tumor cell separation and enrichment kit. The circulating tumor cell separation and enrichment kit comprises a circulating tumor cell separation and enrichment preservation solution, and the circulating tumor cell separation and enrichment preservation solution is an aqueous solution containing the following components by concentration of 3-4 mmol / L ofNaCl, 12-14 g / L of NH4Cl, 11.5-14 mmol / L of Tris-HCl, 29-31.5 mmol / L of 4-hydroxyethyl piperazine ethanesulfonic acid, and 0.03-0.06 w / v% of corn oligopeptides. The circulating tumor cell separation and enrichment preservation solution can effectively preserve CTCs in a biological sample, keep the surface and cell internal antigen activity of the CTCs, improve the enrichment efficiency of the CTCsin the sample to be detected, has a very good erythrocyte lysis effect, and can realize normal-temperature storage or transportation of the CTCs-containing biological sample.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a circulating tumor cell separation and enrichment kit. Background technique [0002] Circulating tumor cells (CTCs) are tumor cells that shed from the primary and metastatic lesions of solid tumors and enter the human peripheral blood circulation. Compared with other normal cells in peripheral blood, circulating tumor cells are derived from tumor tissue, and generally have the characteristics of large cell volume, low density, low rigidity, and high membrane capacitance, and also express epithelial marker proteins and specific proteins of certain derived tissues. Individualized treatment of tumors requires a real-time understanding of the genetic status of tumor cells. CTCs are derived from tumor tissues, and their genetic characteristics are highly consistent with those of tumor tissues, and the changes in the number and type of CTCs are closely related to chan...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01N1/02
CPCC12N5/0693A01N1/0226C12N2509/00
Inventor 许嘉森陈至里黄洁芬吴诗扬
Owner SUREXAM BIO TECH
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