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Cell preservation solution, cell preservation tube and preservation method

A preservation method and technology of preservation solution, applied in the field of medicine, can solve problems such as poor effect and poor preservation effect of circulating tumor cells

Active Publication Date: 2021-07-30
深圳天烁生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Immunomagnetic bead capture and immunofluorescence identification are commonly used methods to enrich and identify circulating tumor cells, but the existing reagents are not effective in preserving circulating tumor cells in peripheral blood, so that circulating tumor cells are enriched by immunomagnetic beads and The effect of further immunofluorescence identification is not good

Method used

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  • Cell preservation solution, cell preservation tube and preservation method
  • Cell preservation solution, cell preservation tube and preservation method
  • Cell preservation solution, cell preservation tube and preservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Preparation of cell preservation solutions with different formulations

[0053] 1. Take KCl, NaCl and Na respectively 2 HPO 4 , adenosine and reduced glutathione, prepare mother liquor, the mother liquor concentration of described KCl is 100mmol / L, and the mother liquor concentration of described NaCl is 1mol / L, and the NaCl 2 HPO 4 The concentration of the mother solution is 1mol / L, the concentration of the mother solution of the adenosine is 100mmol / L, and the concentration of the mother solution of the reduced glutathione is 100mmol / L;

[0054] 2. Mix the KCl, the NaCl, the adenosine, the reduced glutathione and the Na 2 HPO 4 mother liquor, add EDTA-2Na and glucose, dilute with distilled water, stir and mix, and adjust the pH to 7.2;

[0055] 3. Constant volume of the solution;

[0056] 4. Use a filter membrane with a filter hole diameter of 0.2 μm to filter and sterilize the solution in the previous step to obtain a cell preservation solution, and pour th...

Embodiment 2

[0110] Collect 4ml of peripheral blood from 27 patients with lung cancer, add 1ml of the eight groups of cell preservation solution in Example 1, mix upside down, and place at room temperature for 3 days, 5 days and 7 days respectively. Using the immunomagnetic bead capture and immunofluorescence identification methods in Example 1, the preservation effect of the cells was detected. Please refer to Table 3 for the results of cell staining and counting, and the values ​​are mean values.

[0111] Table 3 Example 2 Cell staining and counting results

[0112]

[0113] It can be seen from Table 3 that the cell preservation solution of the present invention can stably preserve the peripheral blood of lung cancer patients for 7 days, and during preservation, the expression of surface antigens of leukocytes and circulating tumor cells (CTCs) can be maintained, which can be captured by immunomagnetic beads and further immunofluorescence Identification. The cell preservation soluti...

Embodiment 3

[0115] Collect 4 ml of peripheral blood from 31 patients with gastric cancer, 20 patients with breast cancer, and 25 patients with liver cancer, add 1 ml of the eight groups of cell preservation solutions in Example 1, mix them upside down, and place them at room temperature for 7 days. Using the immunomagnetic bead capture and immunofluorescence identification methods in Example 1, the preservation effect of the cells was detected. See Table 4 for cell staining count results.

[0116] Table 4 Example three cell staining and counting results

[0117]

[0118]

[0119] It can be seen from Table 4 that the cell preservation solution of the present invention can stably preserve the peripheral blood of gastric cancer patients, breast cancer patients and liver cancer patients for 7 days, and can also maintain the surface of gastric cancer circulating tumor cells, breast cancer circulating tumor cells and liver cancer circulating tumor cells during the preservation period. An...

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Abstract

The invention discloses a cell preservation solution, a cell preservation tube and a preservation method, and the cell preservation solution comprises 2 to 2.5 mmol / L of KCl, 130 to 150 mmol / L of NaCl, 10 to 12 mmol / L of Na2HPO4, 1 to 8 mmol / L of adenosine, 1 to 2 mmol / L of reduced glutathione, 10 to 15 g / L of an anticoagulant and 0.1 to 1 g / L of glucose. The cell preservation solution can stabilize the activity of circulating tumor cells in peripheral blood and the expression of surface antigens of the circulating tumor cells for a long time, so that the effect that the circulating tumor cells are captured by immunomagnetic beads and further subjected to immunofluorescence identification is improved.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a cell preservation solution, a cell preservation tube and a preservation method. Background technique [0002] Cancer cells shed from the primary site and enter the peripheral blood of the human body, which are called circulating tumor cells (Circulating Tumor Cell, CTC). Circulating tumor cells spread to other organs of the body through blood dissemination, which is the main cause of death of patients. Immunomagnetic bead capture and immunofluorescence identification are commonly used methods to enrich and identify circulating tumor cells, but the existing reagents are not effective in preserving circulating tumor cells in peripheral blood, so that circulating tumor cells are enriched by immunomagnetic beads and Further immunofluorescence identification did not work well. [0003] The above content is only used to assist in understanding the technical solution of the inventi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0226A01N1/0205
Inventor 张磊
Owner 深圳天烁生物科技有限公司
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