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

Separation and culture method of ovarian granular cells of northern Guizhou Ma goats

A Qianbei hemp sheep and granular cell technology, applied in the field of cell biology, can solve the problems of few cleaning times, no clear regulation of cleaning times, probability of cell contamination, etc., to improve accuracy, easy operation, and fewer dead cells. Effect

Inactive Publication Date: 2021-02-05
GUIZHOU UNIV
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, in the process of separating ovarian granulosa cells, there is no clear regulation on the number of times of cleaning, whether it is the treatment of the ovary or the post-centrifugation treatment of the cells. number of viable cells

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
  • Separation and culture method of ovarian granular cells of northern Guizhou Ma goats
  • Separation and culture method of ovarian granular cells of northern Guizhou Ma goats
  • Separation and culture method of ovarian granular cells of northern Guizhou Ma goats

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The method for isolating and culturing ovary granulosa cells of Qianbei Ma sheep comprises the following steps:

[0041] (1) The adult purebred Qianbei Ma sheep ewes were sacrificed by neck bleeding, the ovaries and follicles were separated, and 75% alcohol was used to sterilize for 35 minutes at 110°C and 100kPa, and 1:1 bluegrass was added. and streptomycin mixed solution in PBS buffer to remove blood stains, rinsed 3 times and then transferred to a beaker filled with PBS buffer pre-cooled to 4°C, and quickly transferred to a sterile room within 4 hours. During the transfer process, the temperature was kept at 4°C;

[0042] (2) Take out the ovaries and follicles treated in step (1), soak them in 75% alcohol for 30 seconds, and press 75% alcohol-physiological saline-sterilized PBS buffer solution (containing 1:1 penicillin and streptomycin solution) - Sequential washing of DME / F12 medium 3 times;

[0043] (3) Put the ovaries and follicles treated in step (2) in a pet...

Embodiment 2

[0048] The method for isolating and culturing ovary granulosa cells of Qianbei Ma sheep comprises the following steps:

[0049] (1) The adult purebred Qianbei Ma sheep ewes were sacrificed by neck bleeding, the ovaries and follicles were separated, sterilized with 75% alcohol and sterilized at 120°C and 100kPa for 20 minutes, and added 1:1 green and streptomycin mixed solution in PBS buffer to remove blood stains, washed 3 times, then transferred to a beaker filled with PBS buffer pre-cooled to 4°C, and quickly transferred to a sterile room within 5 hours. During the transfer process, the temperature was kept at 8°C;

[0050] (2) Take out the ovaries and follicles treated in step (1), soak them in 75% alcohol for 30 seconds, and press 75% alcohol-physiological saline-sterilized PBS buffer solution (containing 1:1 penicillin and streptomycin solution) - Sequential washing of DME / F12 medium 5 times;

[0051] (3) Put the ovaries and follicles treated in step (2) in a petri dish...

Embodiment 3

[0056] The method for isolating and culturing ovary granulosa cells of Qianbei Ma sheep comprises the following steps:

[0057] (1) The adult purebred Qianbei Ma sheep ewes were sacrificed by neck bleeding, the ovaries and follicles were separated, sterilized with 75% alcohol and sterilized at 115°C and 90kPa for 30 minutes, and added 1:1 green and streptomycin mixed solution in PBS buffer to remove blood stains, washed 3 times, then transferred to a beaker filled with PBS buffer pre-cooled to 4°C, and quickly transferred to a sterile room within 4.8 hours. Keep at 7°C;

[0058] (2) Take out the ovaries and follicles treated in step (1), soak them in 75% alcohol for 30 seconds, and press 75% alcohol-physiological saline-sterilized PBS buffer solution (containing 1:1 penicillin and streptomycin solution) - Sequential washing of DME / F12 medium 5 times;

[0059] (3) Put the ovaries and follicles treated in step (2) in a petri dish filled with 100ml of 10% complete medium, where...

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 present invention discloses a separation and culture method of ovarian granular cells of northern Guizhou Ma goats. The separation and culture method of ovarian granular cells of northern GuizhouMa goats comprises the following steps: sacrificing adult multiparous purebred northern Guizhou Ma goats by neck bloodletting, taking out ovaries, flushing away bloodstain, bringing the treated ovaries back to a sterile room, performing soaking with alcohol, rinsing with alcohol, normal saline and sterilized PBS, puncturing follicles with a needle, releasing ovarian granular cells, performing rinsing with a 10% complete medium, after centrifuging, taking a precipitate, continuously rinsing the precipitate with a sterilized PBS buffer solution, adding a 10% complete medium, and peforming blowing, beating, uniformly mixing, and culturing in a constant-temperature incubator. The pure ovarian granular cells can be separated through the method and the method is easy and convenient to operate, stable, and reliable and has good repeatability.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for separating and culturing ovarian granulosa cells of Qianbei Ma sheep. Background technique [0002] Qianbei Ma sheep is an excellent local goat breed in Guizhou. It has the advantages of tolerance to rough feeding, strong disease resistance, docile sex, good meat quality, good quality of lamb skin, and higher reproductive rate than other local breeds in Guizhou. It is an excellent material for breeding multi-lamb strains. [0003] As the main functional cells of the ovary, granulosa cells play an important role in the whole development process from the generation of primordial follicles to the discharge of mature follicles. Granulosa cells can act as a medium to make up for the lack of oocyte uptake of small molecules through gap junctions. At the same time, the growth and differentiation of granulosa cells is the key to the initiation and growth of primordial follicles....

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): C12N5/071
CPCC12N5/0682C12N2509/00
Inventor 陈祥敖叶周志楠洪磊唐文杨沛方
Owner GUIZHOU UNIV
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com