Preparation method and application of bladder urothelial single cell suspension for single cell sequencing

A technology of single-cell sequencing and single-cell suspension, which is applied in the field of biomedicine, can solve problems such as inappropriateness, and achieve the effect of easy operation and good practical application value

Active Publication Date: 2021-04-02
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since single-cell sequencing requires a large number of isolated and highly active cells (survival rate above 90%), most current methods for studying urothelial cells, whether isolating fresh cells or obtaining primary cultured cells, are not feasible. Ideal for single-cell sequencing

Method used

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  • Preparation method and application of bladder urothelial single cell suspension for single cell sequencing
  • Preparation method and application of bladder urothelial single cell suspension for single cell sequencing
  • Preparation method and application of bladder urothelial single cell suspension for single cell sequencing

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Experimental program
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Effect test

Embodiment

[0049] animal

[0050] Mice (8-10 weeks old) were purchased from the Animal Experiment Center of Shandong University. All mice were kept in the institute's specific pathogen-free animal facility.

[0051] Reagent

[0052] D-PBS solution (without calcium, magnesium, phenol red) (Solarbio, Chinacat.no.D1040)

[0053] BSA (Solarbio, China cat. no. A8850)

[0054] DMEM medium (Thermo Fisher Scientific, USA cat.no.C11995500BT)

[0055] PBS-EDTA solution (10mmol / L) (leagene, China cat.no.CZ0026)

[0056] Dispase II (Sigma, USA cat. no. 04942078001)

[0057] TrypLE TM Express Enzyme (1X, no phenol red) (Thermo Fisher Scientific, USA cat. no. 12604-013)

[0058] Sylgard 184 Silicone Elastomer Kit (Dow Corning, USA cat.no.DC184)

[0059] 75% ethanol

[0060] equipment

[0061] Curved hemostat for opening the chest cavity (Yuyan, China cat.no.Y30373)

[0062] Tissue scissors for opening the thorax and abdomen (Yuyan, China cat.no.Y2201)

[0063] Two pairs of dissecting scis...

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Abstract

The invention provides a production method of a bladder urothelium single-cell suspension used for single-cell sequencing, and belongs to the technical field of biomedicine. The production method comprises conducting systemic hemoperfusion on sacrificed animals, and conducting incubation and enzymolysis digestion on collected bladders to obtain the single-cell suspension. By means of the bladder urothelium single-cell suspension produced through the production method of the bladder urothelium single-cell suspension used for single-cell sequencing, not only is the cell quality guaranteed, but also a large number of separated urothelium cells can be obtained at once; and the method does not require highly specialised equipment, reagents or skills, and operation of researchers is facilitated,so that the method has good practical application values.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a preparation method and application of bladder urothelial single-cell suspension for single-cell sequencing. Background technique [0002] The information disclosed in the Background of the Invention is only intended to increase the understanding of the general background of the invention, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] The mammalian bladder is like a hollow sphere composed of the urothelium, lamina propria, detrusor muscle, and serosa ( figure 1 A). The urothelium is a special layered epithelium composed of at least three cell types: umbrella cells, intermediate cells, and basal cells ( figure 1 B). These cells act as a barrier to the urinary tract by generating and maintaining crystalline plaques. Diff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/02
CPCC12N5/0685C12N2509/00
Inventor 史本康刘亚骁李岩
Owner SHANDONG UNIV QILU HOSPITAL
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