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Method for obtaining sheep endometrial epithelial cells and stroma cells, separation and purification method and application

A technology for endometrial and epithelial cells, applied in the biological field, can solve problems such as unfavorable cell proliferation, inability to passage, and inability to grow for a long time.

Inactive Publication Date: 2020-09-22
SHIHEZI UNIVERSITY
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  • Abstract
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Problems solved by technology

The results showed that tissue cultured cow and sheep endometrial cells grew faster, and the cell morphology was spindle-shaped and paving stone-like intertwined; normal temperature digestion method did not succeed in obtaining cow endometrial cells, but obtained sheep endometrial cells ; while the first tissue culture and then digestion method obtained dairy cow endometrial cells, but they could not be passaged and could not grow for a long time; while the cells obtained by first digestion and then tissue culture were similar to the cow and sheep endometrial cells obtained by single group culture , there are many tissue fragments on the cells, covering the cells is not conducive to cell proliferation

Method used

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  • Method for obtaining sheep endometrial epithelial cells and stroma cells, separation and purification method and application
  • Method for obtaining sheep endometrial epithelial cells and stroma cells, separation and purification method and application
  • Method for obtaining sheep endometrial epithelial cells and stroma cells, separation and purification method and application

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Embodiment

[0037] (1) Rinse the sheep uterus obtained from the slaughterhouse with PBS containing 5% penicillin-streptomycin double antibody for 3 times, cut off 2 / 3 of the uterine horns under aseptic conditions and place them in a disposable petri dish. After being cut longitudinally, unfold the caruncles on the surface of the uterine cavity to expose the caruncles and cut off the caruncles with scissors. PBS (5% double antibody) washed the excised caruncle 3 times.

[0038] (2) Put the uterine caruncle into a 2mL centrifuge tube with tweezers, and cut the uterine caruncle into 1mm pieces with scissors 3 Small pieces, washed with PBS.

[0039] (3) Use a pipette gun to inoculate 500 μL of shredded tissue pieces into a 6-well plate, add 2 mL of D-MEM / F-12 (containing 10% FBS, 2% double antibody) and place at 37°C, 5% CO 2 Culture in a medium incubator, and then change the medium every 3 days. On day 3 the tissue pieces began to climb out of the cells (eg figure 1 ), the cells reached ...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for obtaining sheep endometrial epithelial cells and stroma cells, a separation and purification method and an application. A large amount of the sheep endometrial epithelial cells and stroma cells are simply, conveniently and rapidly obtained by using a tissue block culture method, and separation parts are changed. According to the method, uterine horns are longitudinally cut open, round uterine cotyledons on the uterine horns are sheared off after the uterus horns are unfolded, the uterine cotyledons are clamped into a 2 mL centrifuge tube by tweezers, and then the uterine cotyledons are sheared into 1 mm<3> small blocks in the centrifuge tube by small scissors. The method is simpler and more convenient. Meanwhile, the epithelial cells and the stroma cells are purified through time difference digestion in the method. Due to different tolerance of the stroma cells and the epithelial cells totrypsin, the stromal cells are easier and faster to digest, and purification is carried out by utilizing time difference. The method is simpler and easier to master than an existing digestion and sieving technology.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for obtaining sheep endometrium epithelial cells and stromal cells, a separation and purification method and application. Background technique [0002] Endometrial epithelial cells (endometrial epithelium cells, EECs) and endometrial stromal cells (endometrial stromal cells, ESCs) are the main constituent cells of the endometrium, both of which are affected by ovarian hormones to produce periodic changes. In order to better study the response between the uterus and the embryo, the endometrium is isolated and the primary cell culture is carried out in vitro, which can establish a model for the in vitro study of the influence of the endometrium on the embryo and create conditions for the study of cell structure and function. . [0003] There are many methods of primary culture, the most basic and commonly used are two kinds, namely tissue block method and digestio...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2509/00
Inventor 胡广东王静凌芳郝科兴谷新利曾维斌黄涛陈慧慧龙德智
Owner SHIHEZI UNIVERSITY
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