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Preparation method of human liver cancer tissue and liver tissue active single cell suspension

A single-cell suspension and liver cancer tissue technology, applied in the biological field, can solve the problem of low success rate of single-cell suspension preparation, and achieve the effect of simple operation, good stability and high cell activity

Pending Publication Date: 2020-08-25
SUN YAT SEN UNIV CANCER CENT
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is mainly used to solve the problem of low success rate of single cell suspension preparation when performing single cell transcriptome sequencing on liver tissue and liver cancer tissue

Method used

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  • Preparation method of human liver cancer tissue and liver tissue active single cell suspension
  • Preparation method of human liver cancer tissue and liver tissue active single cell suspension
  • Preparation method of human liver cancer tissue and liver tissue active single cell suspension

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Step 1. Storage and transfer of tissue samples:

[0043] Prepare a plastic ice box about 40cm x 30cm x 40cm in length, width and height, with appropriate amount of small ice cubes inside. 2. Take a number of 15ml plastic sampling cups (or 15ml centrifuge tubes), and add 5-8ml of serum-free medium DMEM or 1640, which has been pre-cooled in a refrigerator at 4°C, to them. 3. Quickly soak the punctured tissue specimens into the container Put the sampling cup (or centrifuge tube) into the sampling cup (or centrifuge tube) with medium, and then insert the sampling cup (or centrifuge tube) into a plastic ice box for storage and transport. 4. The shorter the transit time, the better, try to control the transit time within 20 minutes.

[0044] Step 2. Device material preparation:

[0045] Prepare a clean 6-well plate, take 3 wells, add 5ml pre-cooled (4°C) serum-free medium (DMEM or 1640) to 2 wells, add 5ml serum-free culture + three digestive enzymes to 1 well ( A, H, R). ...

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PUM

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Abstract

The invention provides a preparation method of human liver cancer tissue and liver tissue active single cell suspension. The preparation method includes the following steps: S1, preserving and transporting tissue samples; S2, performing rinsing and pre-digestion on the tissue; S3, performing tissue digestion; S4, terminating digestion, performing filtering and removing impurities; S5, cracking redblood cells, and performing secondary filtering; S6, washing and purifying cells, and removing necrotic cells; and S7, performing re-suspension to obtain the cells. The cell activity of the single cells obtained by the preparation method is good, and the total activity of the cells can be greater than 70%; and the preparation method is simple in operation and good in stability.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of human liver cancer tissue and active single cell suspension of liver tissue. Background technique [0002] In recent years, biological research has made great progress in the field of single cells. Technical platforms such as Cytof mass spectrometry, BDRhapsody, and 10×Genomics emerged as the times require. At present, the most popular research is single-cell transcriptome sequencing, which can analyze in detail the specific differences between different types of cells in complex samples to a certain extent, and obtain some key data that were previously covered up by whole-transcriptome sequencing. In addition, related research on single-cell proteomics is also being explored. Although single-cell quantitative detection of selected protein combinations is currently only possible, single-cell protein quantification can also obtain differences betwe...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/071
CPCC12N5/0693C12N5/067C12N2509/00
Inventor 王小辉徐立陈敏山王骏成施木德张耀军邹如海
Owner SUN YAT SEN UNIV CANCER CENT
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