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

Method for separating rat testis interstitial Leydig cells

A testicular interstitial, cell technology, applied in the field of biomedicine, can solve the problems of low cell activity, time-consuming, low cell recovery rate, etc., and achieve the effect of low cost

Pending Publication Date: 2022-01-25
THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although these currently available separation methods for LCs are able to obtain high-purity cells (about 95%), they either have low cell recovery or low cell viability or are time-consuming, and more importantly, these methods require special equipment such as panning. Washing instrument or flow cytometer with sorting capability

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
  • Method for separating rat testis interstitial Leydig cells
  • Method for separating rat testis interstitial Leydig cells
  • Method for separating rat testis interstitial Leydig cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] A method for separating Leydig cells of rat testis, comprising the steps of:

[0040] Obtain rat testis and prepare rat testis cell suspension;

[0041] Labeled rat Leydig cells;

[0042] The positive cells were screened by magnetic bead sorting (MACS);

[0043] Positive cells were purified repeatedly by magnetic bead sorting.

[0044]Firstly, the rat testis is prepared into a cell suspension, and the preparation method of the suspension is mainly to digest the parenchymal tissue of the testis in a culture medium; the digested tissue suspension is allowed to stand at room temperature, and the supernatant is filtered to obtain Testicular cell suspension for easy separation. Since one of the innovations of this method is to use the magnetic bead sorting method to screen rat Leydig cells, it is necessary to cooperate with magnetic markers to mark the magnetic particle markers on the surface of rat Leydig cells, and then perform positive cell selection. Transfer the mag...

Embodiment 2

[0060] Reagent

[0061] DMEM / F12 cell culture medium, bovine serum albumin (BSA), collagenase type IV, testosterone, testosterone-d3, and penicillin / streptomycin double antibodies were purchased from Sigma-Aldrich (St. Louis, Missouri). Trypan blue solution (0.4%) was from ThermoFisher (Waltham, MA). The reverse transcription kit was from Promega (Madison, WI). 480 Green I Master is from Roche (Basel, Switzerland). Anti-mouse IgG1 magnetic particles and BD IMag TM Buffer (10X) was from BD Biosciences (Franklin Lakes, NJ). LH was from MyBiosource (San Diego, CA). Dihydrotestosterone (DHT) and 5α-androstane, 3α, 17α-diol (3α-diol) and 25-hydroxy-vitamin D (25-OH-VD) ELISA kits were purchased from Jianglai Biotechnology Technology Co., Ltd. (Shanghai, China). Dexmedetomidine (DEX) was from Yangzijiang Pharmaceutical Group Co., Ltd. (Taizhou, China). Acetonitrile and methanol were from Merck (NJ, NJ).

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 method for separating rat testis interstitial Leydig cells. The method comprises the following steps: acquiring a rat testis and preparing a rat testis cell suspension; marking rat testicular cells; screening positive cells by adopting a magnetic bead sorting (MACS) method; and repeatedly purifying the positive cells through adoption of a magnetic bead sorting method. The separation method disclosed by the invention is simple and easy to operate, high in cell yield, high in purity and high in activity, and does not need expensive equipment.

Description

technical field [0001] The invention relates to biomedicine, in particular to a method for separating Leydig cells of rat testes. Background technique [0002] The main function of Leydig cells (LCs) in the testis is to produce testosterone (T), a male hormone essential for reproductive function and the development and maintenance of male characteristics. Defects in the number or function of LCs can lead to complex symptoms including changes in body composition, increased fatigue, sexual dysfunction, depressed mood, decreased cognitive function, and reduced immune response. Due to their critical role in male reproduction and general well-being, LCs have been intensively studied over the years in both in vivo and in vitro settings, including in the field of toxicology. In vitro studies of LCs require isolating and culturing cells under the best possible conditions, with minimal interference from other testicular cells, in order to examine cell function under optimal physiolo...

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): C12N5/071C12N13/00
CPCC12N5/0683C12N13/00C12N2509/00C12N2529/00
Inventor 陈浩林赵星仪季敏鹏温馨陈丹
Owner THE SECOND HOSPITAL AFFILIATED TO WENZHOU MEDICAL COLLEGE
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