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Method for separating and cultivating endometrial stem cells of menstrual blood sources

A technology of separation and culture and blood source palace, which is applied in the field of stem cells, can solve the problems of small number of stem cells, low differentiation potential, and contaminated menstrual blood, so as to achieve the effect of rich sources, avoiding menstrual blood pollution, and avoiding external pollution

Inactive Publication Date: 2016-06-01
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, for the isolation and culture of endometrial stem cells, most of them are directly obtained, which causes great harm to patients, and at the same time, the source is suffering and is subject to ethical restrictions. Although there is a small amount of research on menstrual blood-derived stem cells, there is no simple and convenient method. Collection of menstrual blood leads to a higher probability of menstrual blood contamination, fewer stem cells obtained, lower purity, lower differentiation potential, higher cost and complicated process, etc.

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  • Method for separating and cultivating endometrial stem cells of menstrual blood sources
  • Method for separating and cultivating endometrial stem cells of menstrual blood sources
  • Method for separating and cultivating endometrial stem cells of menstrual blood sources

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Embodiment approach

[0037] Concrete embodiments of the present invention are as follows:

[0038] 1. Preparation of menstrual blood collection kit:

[0039] Including a sterile specimen preservation bottle (containing 5-15ml of menstrual blood preservation solution), a sterilized menstrual cup, a pair of gloves, a Pasteur pipette, and an instruction manual.

[0040] The menstrual blood preservation solution contains (2-5)×penicillin, (2-5)×streptomycin, 30-50μg / ml kanamycin, 30-50μg / ml sodium citrate, 100U / ml heparin sodium Phosphate buffer.

[0041] 2. Collection of menstrual blood

[0042] After consultation and consent, collect 5-10ml of menstrual blood samples from healthy women (20-40 years old) according to the instructions of the menstrual blood collection kit, put them in a sterile specimen preservation bottle, mix them thoroughly, store them at low temperature, and process them within 24 hours.

[0043] 3. Sterility test:

[0044] Take a sample of 0.5-1ml under sterile conditions and...

Embodiment 1

[0057] 1. Preparation of menstrual blood collection kit:

[0058] Including a sterile specimen preservation bottle (containing 5ml of menstrual blood preservation solution), a sterilized menstrual cup, a pair of gloves, a Pasteur pipette, and an instruction manual.

[0059] The menstrual blood preservation solution is a phosphate buffer solution containing 2×penicillin, 5×streptomycin, 40 μg / ml kanamycin, 50 μg / ml sodium citrate, and 100 U / ml heparin sodium.

[0060] 2. Collection of menstrual blood

[0061] After consultation and consent, collect 5-10ml of menstrual blood samples from healthy women (20-40 years old) according to the instructions of the menstrual blood collection kit, put them in a sterile specimen preservation bottle, mix them thoroughly, store them at low temperature, and process them within 24 hours.

[0062] 3. Sterility test:

[0063] Under sterile conditions, take 0.5-1ml of a sample and use a rapid microbial detector to detect whether there is bacteri...

Embodiment 2

[0074] 1. Preparation of menstrual blood collection kit:

[0075] Including a sterile specimen preservation bottle (containing 15ml of menstrual blood preservation solution), a sterilized menstrual cup, a pair of gloves, a Pasteur pipette, and an instruction manual.

[0076] The menstrual blood preservation solution is a phosphate buffer solution containing 3×penicillin, 4×streptomycin, 50 μg / ml kanamycin, 30 μg / ml sodium citrate, and 100 U / ml heparin sodium.

[0077] 2. Collection of menstrual blood

[0078] After consultation and consent, collect 5-10ml of menstrual blood samples from healthy women (20-40 years old) according to the instructions of the menstrual blood collection kit, put them in a sterile specimen preservation bottle, mix them thoroughly, store them at low temperature, and process them within 24 hours.

[0079] 3. Sterility test:

[0080] Under sterile conditions, take 0.5-1ml of a sample and use a rapid microbial detector to detect whether there is bacter...

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Abstract

The invention relates to the field of stem cells, in particular to a method for separating and cultivating endometrial stem cells of menstrual blood sources. The method has the advantages that female menstrual blood is used as a stem cell acquisition source, accordingly, waste can be utilized and can be turned into wealth, and the stem cell acquisition source is abundant; the quantities of the abundant endometrial stem cells in the menstrual blood are 30 times the contents of effective constituents in bone marrow, and accordingly the endometrial stem cells are high in content; the menstrual blood is preserved by the aid of menstrual blood preserving fluid and accordingly can be prevented from being contaminated by the outside in procedures from acquisition to separation; the endometrial stem cells are separated by the aid of interlayer centrifugal tubes and cell adherence processes which are combined with one another, and are cultivated by the aid of serum-free media, accordingly, the method is easy to implement and low in cost, and large quantities of endometrial stem cells with high purity and differentiation capacity can be obtained from the menstrual blood.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a method for isolating and culturing endometrial stem cells derived from menstrual blood. Background technique [0002] The uterus is a tissue with high regenerative and differentiation capabilities. In each menstrual cycle, the endometrium of a woman can grow from about 0.5mm to about 5-7mm. A woman can experience 400 such cycles in her lifetime. Studies have shown that the endometrium contains stem cells with high differentiation potential, proliferative ability and low immunogenicity. However, its direct acquisition methods, including hysterectomy, early pregnancy hysterectomy, and curettage, are all severely invasive to the donor. [0003] In recent years, Japanese scientists have discovered stem cells with multi-directional differentiation potential in women's menstrual blood, which can be used to repair damaged myocardial tissue. In 2008, American scientists Patel et al. discove...

Claims

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Application Information

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IPC IPC(8): C12N5/074
CPCC12N5/0682C12N2509/00C12N2509/10
Inventor 陈海佳王一飞葛啸虎马岩岩王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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