Single cell separation method of Aurelia coerulea polyps

Abstract

The invention provides a single cell separation method of Aurelia coerulea polyps. The method comprises the following steps of A, preparing artificial seawater; B, preparing a polyp sample, specifically, transferring polyps into a culture container filled with clean sterile artificial seawater, and placing the culture container in an incubator at 20 DEG C; hungering for 24 hours before use, selecting polyps with stretched tentacles for later use, and cutting the sample into small fragmented blocks; and C, preparing single cells, specifically, preparing a 30g / L sodium chloride solution, and preparing a dissociation enzyme solution 1 or 2 by using the sodium chloride solution as a solvent; adding the small sample fragmented blocks into a dissociation enzyme solution for digestion to obtain a digestion mixed solution; and filtering the mixed solution through a cell sieve, centrifuging the filtrate at 4 DEG C, repeatedly cleaning the precipitate with a dissociation solution, and re-dissolving in the dissociation solution to obtain a single-cell suspension, wherein the dissociation enzyme solution 1 comprises 0.01 to 1.5 g / L of collagenase I, 0.01 to 1 g / L of neutral protease and 0.01 to 0.5 g / L of elastase; and the dissociation enzyme solution 2 comprises 0.01 to 2 g / L of collagenase IV and 0.01 to 20 U / mL of papain.

Description

technical field [0001] The invention belongs to the technical field of single cells, and in particular relates to a method for separating single cells of the hydra of sea moon jellyfish. Background technique [0002] With the rapid development of technology, DNA and RNA sequencing have become the most commonly used scientific research methods. People use sequencing to detect all transcripts in samples or carry out gene expression analysis. The objects of this method of sequencing are often tissue samples or cells. clusters, which makes it possible for differences between cells to be masked by the mean. Single-cell transcriptome sequencing solves this problem very well. It can capture a single cell, thereby independently providing the RNA expression profile of each cell, and effectively revealing the differences between cells. [0003] The moon jellyfish (Aurelia coerulea) belongs to the phylum Cnidaria, the class Aurelia, the order Aurelia, the family Aureliaceae, and the g...

AI Technical Summary

Problems solved by technology

However, the existing single cell separation method is only suitable for the separation of single cells of freshwater organisms, coupled with the characteristics of easy autolysis of polyps, the resulting single cell viability is low, which limits the development of polyp cell level research

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