Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation method of fish tissue single cell suspension

A single-cell suspension, fish technology, applied in the field of gene detection, can solve the problems of inability to obtain cell viability tissue single-cell suspension, restricting the establishment of cell culture cell lines and functional gene verification research work, etc. Good viability and high efficiency

Pending Publication Date: 2021-09-17
SHANGHAI PASSION BIOTECHNOLOGY CO LTD
View PDF7 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing experimental techniques are still unable to obtain tissue single-cell suspensions with good cell viability, which restricts research work on tissue cell culture, establishment of cell lines, and functional gene verification.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation method of fish tissue single cell suspension
  • Preparation method of fish tissue single cell suspension
  • Preparation method of fish tissue single cell suspension

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0029] A preparation method of fish tissue single cell suspension, comprising the following steps:

[0030] 1) Preparation of reagents and culture medium, including serum-free medium and complete medium, preparation of cell cleaning solution, preparation of mixed enzyme dissociation stock solution;

[0031] 2) Tissue pretreatment, add an appropriate amount of 1*DPBS solution in a sterile RNase-free cell culture dish; cut the tissue into 0.5-1mm 2 After small pieces, inhale into a 15ml test tube containing 1*DPBS solution, and rinse with 1*DPBS solution for 1-2 times until the excess tissue is removed;

[0032] 3) Digest and dissociate the tissue, suck off excess 1*DPBS solution, add 3-5ml mixed enzyme dissociation stock solution; digest and incubate in a water bath at 37°C for 15 minutes;

[0033] 4) Termination of digestion: Add an equal volume of culture medium solution prepared earlier, gently pipette and mix well, no small tissue segments can be seen with the naked eye, a...

Embodiment 1

[0049] Step 1: Configure reagents and media

[0050] 1. Prepare the following two media respectively

[0051] (1) Serum-free medium: the aliquoted M199-HEPES medium contains 1% Penicillin-Streptomycin; add 1ml Penicillin-Streptomycin to 99ml M199-HEPES medium

[0052] (2) Complete medium: the aliquoted M199-HEPES medium contains 10% FBS and 1% Penicillin-Streptomycin; 89ml M199-HEPES medium is added with 10ml FBS and 1ml Penicillin-Streptomycin;

[0053] 2. Prepare cell washing solution: HBSS-5% FBS solution: mix 95ml HBSS solution with 5ml lFBS solution;

[0054] 3. Prepare mixed enzyme dissociation stock solution: Dissolve 10mg type II collagenase lyophilized powder (Sigma, product number C6885-1G), 5mg DispaseII lyophilized powder (Sigma, product number D4693-1G) in 10ml serum-free M199-HEPES culture (Sigma, Cat. No. 12340030), after being completely dissolved, filter-sterilize and store in a -20°C freezer to avoid repeated freezing and thawing.

[0055] Step Two: Tissue...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a preparation method of a fish tissue single cell suspension. The preparation method comprises the following steps of: 1) preparing a reagent and a culture medium; 2) pretreating tissues; 3) performing tissue digestion and dissociation; 4) terminating digestion; 5) filtering by using a micron cell filter; 6) obtaining a target tissue single cell suspension; and 7) measuring the yield of living cells. The preparation method of the fish tissue single cell suspension can be used for quickly and efficiently obtaining the fish tissue single cell suspension at low cost, ensures that dissociated cells keep good survival rate, can be applied to dissociation, separation and purification of large-scale fish cells and establishment of cell lines, and provides important reference data for research of fish single cells, gene functions and genetic breeding.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method for preparing fish tissue single-cell suspension. Background technique [0002] In recent years, with the rapid development of technology in the field of biological research, single-cell sequencing technology has emerged as the times require. technology. Single-cell sequencing can reveal the gene structure and gene expression of a single cell, classify and compare cells, and reflect the heterogeneity between cells. It plays an important role in the fields of tumors, developmental biology, immunology, and microbiology. [0003] Different tissue cells have their own unique physiological characteristics, which is the main reason why tissue cell culture has become a worldwide problem. Existing experimental techniques are still unable to obtain tissue single-cell suspensions with good cell viability, which restricts research work on tissue cell culture, establishment...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/071C12Q1/6869
CPCC12N5/0602C12Q1/6869C12N2509/00C12N2509/10
Inventor 李靖贇余婧孙子奎
Owner SHANGHAI PASSION BIOTECHNOLOGY CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com