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Human endometrial stromal cell separation and primary culture method

A technology for endometrium and primary culture, which is applied in the field of human cell separation and culture, and can solve the problems of high technical requirements for primary cell culture, low success rate of endometriotic stromal cell culture, and low success rate

Inactive Publication Date: 2018-11-16
THE FIRST AFFILIATED HOSPITAL OF GUANGXI MEDICAL UNIV +2
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, primary cell culture requires high technology and low success rate, especially the success rate of cultured endometriotic stromal cells is not high. The earliest report by Ryan et al. was that the cultured success rate of endometriotic stromal cells was 56%. In recent years, the success rate reported by relevant scholars is only 60-70%.

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  • Human endometrial stromal cell separation and primary culture method
  • Human endometrial stromal cell separation and primary culture method
  • Human endometrial stromal cell separation and primary culture method

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Embodiment Construction

[0029] How to implement the present invention is illustrated by the following examples, and reagents and equipment used are as follows:

[0030] Main test reagent

[0031]

[0032] Main equipment

[0033]

[0034] 1. Isolation of primary endometrial stromal cells

[0035] 1.1 Solution preparation

[0036] Complete medium: 1640 culture medium + 20% FBS (volume percentage) + 1% double antibody. In view of the tumor-like growth characteristics of heterotopic mesenchymal cells, the present invention selects the 1640 medium commonly used in tumor cell culture to replace the medium reported in conventional literature for the first time, and increases the serum concentration of the culture solution to 20%.

[0037] Mouse anti-human vimentin monoclonal antibody (vimentin): Diluted with PBS solution at a volume ratio of 1:100 and stored at 4°C for a short period of time.

[0038] Mouse anti-human keratin monoclonal antibody: diluted with PBS solution at a volume ratio of 1:20...

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Abstract

The invention discloses a human endometrial stromal cell separation and primary culture method which comprises the following steps of digesting human endometrial tissues by adopting pancreatin withoutcontaining EDTA; during screen filtering of the tissues after digestion, poking filtering tissues to enable the filtering tissues to perform full filtering; and adopting a 1640 culture medium to be used for culture of primary endometrial stromal cells. Compared with the existing method, the method greatly increases the acquisition amount of the primary endometrial stromal cells and the success rate of cell isolated culture, is more efficient and economic and provides a stable and reliable in-vitro cell model to clinical study of endometrial diseases and development of drugs. In addition, an inventor also observes and compares differences between normal endometrial stromal cells and ectopic endometrial stromal cells in the aspects of cellular morphology, the growth characteristic, the biological behavior and the like.

Description

technical field [0001] The invention belongs to the technical field of human cell separation and culture, and in particular relates to a method for the separation and primary culture of human endometrial stromal cells. Background technique [0002] Endometriosis (Ems) is a benign and common gynecological disease. Ovarian endometriosis frequently occurs in gynecological clinics and seriously affects the quality of life of patients. In order to strengthen the basic research on its etiology and pathogenesis, In order to provide a better method for clinical treatment, researchers have been exploring the establishment of in vivo and in vitro experimental models that are conducive to the study of the disease. In vitro models of endometriosis include tissue models and cell models. The tissue model is mainly used to observe the early ectopic tissue invasion, proliferation and angiogenesis process, but the sources and conditions of in vitro culture are different, the endometrial tis...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0652C12N2509/00
Inventor 庞丽红刘俐伶韦懿芸邓翎洁莫琳吴福智
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGXI MEDICAL UNIV
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