Method for separating, culturing and purifying endometrial cells

A technology of endometrium and purification method, which is applied in the direction of cell dissociation method, cell culture active agent, biochemical equipment and method, etc., which can solve the problems of small number of adherent cells, long replacement cycle, decreased cell activity, etc., and achieve The effect of improving the efficiency of isolation and culture

Inactive Publication Date: 2020-04-21
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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  • Application Information

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Problems solved by technology

[0004] The existing separation technology for endometrial cells usually uses 0.25% trypsin to digest for 30 minutes, and then passes through a sieve to obtain a single-cell suspension. serious decline

Method used

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  • Method for separating, culturing and purifying endometrial cells
  • Method for separating, culturing and purifying endometrial cells

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Embodiment

[0030] A method for separating, culturing and purifying endometrial cells, comprising the following steps:

[0031] 1. Take the exfoliated endometrial tissue, add cleaning solution to clean it, add red blood cell lysate to the cleaning solution to remove blood, cut it into pieces with straight scissors, and set aside;

[0032] Wherein, the cleaning solution is prepared by physiological saline containing 50% erythrocyte lysate.

[0033] 2. Take the shredded endometrial tissue in step 1, add digestion solution I, digest at 37°C for 1.5 hours, stop digestion with endometrial cell culture medium after digestion, then filter through a 400-mesh filter, separate and retain the filtrate and filter residue, and centrifuge the filtrate: the centrifugal force is 300g, the time is 5min, and the precipitate obtained after centrifugation is resuspended with endometrial cell culture medium;

[0034] Wherein, the digestive solution I was prepared by adding 3 mg / mL type I collagenase and 0.3 ...

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Abstract

The invention relates to a method for separating, culturing and purifying endometrial cells, and belongs to the technical field of human cell separation and culture. The method comprises the followingsteps: taking an exfoliated endometrial tissue, adding a cleaning solution for cleaning, and cutting the endometrial tissue into pieces for later use after cleaning is completed; adding digestive juice I containing type I collagenase into the cut endometrial tissue, stopping digestion by using an endometrial cell culture solution after digestion is finished, carrying out filtering by using a 400-mesh filter screen, separating filtrate from filter residues, centrifuging the filtrate, and re-suspending the precipitate by using an endometrial cell culture solution to obtain a suspension; and adding trypsin-containing digestive juice II into the filter residues for digestion, stopping digestion by using an endometrial cell culture solution, carrying out centrifuging, taking precipitate, re-suspending the precipitate by using an endometrial cell culture solution to obtain a suspension, and finally, carrying out mixed culture on the suspension and the suspension obtained in the previous step. According to the method for separating, culturing and purifying endometrial cells, more endometrial cells can be obtained more efficiently aiming at the defects of extraction by a conventional method.

Description

technical field [0001] The invention relates to the technical field of separation and cultivation of human cells, in particular to a method for separation, cultivation and purification of endometrial cells. Background technique [0002] The endometrium refers to the inner wall that constitutes the mammalian uterus, and is divided into a functional layer and a basal layer. The endometrium covers the mucous membrane and is composed of the mucosal epithelium and the underlying lamina propria. When estrogen is secreted, each epithelial cell will grow and divide, increasing the number. The portion of the lamina propria below the mucosal epithelium is called the functional layer, and epithelial cells enter the uterus to form the uterine glands and respond to estrogen. [0003] Endometriosis is a common gynecological disease in women caused by active endometrial cells growing outside the endometrium. Endometrial cells should normally grow in the uterine cavity, but because the ut...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/38C12N2501/125C12N2509/00
Inventor 王刚陈扬平赵蓝李静静孙灿兴
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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