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Endometrial stem cell isolation method

An endometrial and separation method technology, applied in the field of stem cell separation, can solve the problem of less separation of exfoliated endometrial-derived cells, and achieve the effects of satisfying the number of clinically used cells, increasing the success rate, and increasing the amount of cells

Active Publication Date: 2019-01-29
BEIJING TAIDONG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, more blood-derived endometrial stem cells are isolated and cultured, and exfoliated endometrium-derived cells are less isolated

Method used

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  • Endometrial stem cell isolation method
  • Endometrial stem cell isolation method
  • Endometrial stem cell isolation method

Examples

Experimental program
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Effect test

Embodiment 1

[0042] 1. Related materials

[0043] 1) Preparation of soluble amphotericin B (effective concentration: 2.5 μg / ml): Add 25 mg of amphotericin B into 5 ml of sterile water, fully dissolve, mix well, and then sterilize through a 0.22 μm filter membrane to prepare Make a stock solution of soluble amphotericin B and store it at -20°C for later use.

[0044] 2) Complete medium: DMEM+10% FBS+2mM glutamine+1% double antibody+2.5μg / ml amphotericin B.

[0045] 3) PBS buffer: add 250 μl soluble amphotericin B stock solution and 15ml double antibody to 500ml PBS, wherein, the double antibody is a mixture of penicillin and streptomycin, the content of penicillin is 10000U / ml, and the content of streptomycin 10mg / ml.

[0046] 2. The method for isolating endometrial stem cells, the steps are as follows:

[0047] Prepare the tweezers and scissors used in the experiment, prepare three Petri dishes, and add 10ml of PBS to the dishes.

[0048] 1) Transfer the collected sample to a 50ml cent...

Embodiment 2

[0069] 1. Related materials

[0070] 1) Preparation of soluble amphotericin B (effective concentration: 2.5 μg / ml): Add 25 mg of amphotericin B into 5 ml of sterile water, fully dissolve, mix well, and then sterilize through a 0.22 μm filter membrane to prepare Make a stock solution of soluble amphotericin B and store it at -20°C for later use.

[0071] 2) Complete medium: DMEM+10% FBS+2mM glutamine+1% double antibody+2.5μg / ml amphotericin B.

[0072] 3) PBS buffer: add 250 μl soluble amphotericin B stock solution and 15ml double antibody to 500ml PBS, wherein the double antibody is a mixture of penicillin and streptomycin, the content of penicillin is 10000U / ml, and the content of streptomycin 10mg / ml.

[0073] 2. The method for isolating endometrial stem cells, the steps are as follows:

[0074] Prepare the tweezers and scissors used in the experiment, prepare three Petri dishes, and add 10ml of PBS to the dishes.

[0075] 1) Transfer the collected sample into a 50ml cen...

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Abstract

The invention relates to the field of separation of stem cells, in particular to an endometrial stem cell isolation method, comprising the following steps: after blood plasma is removed from a sample,conducting diluting, conducting filtering by a sieve of not more than 100 [mu]m, adding antibiotics into an obtained filtrate, then conducting low-temperature treatment, conducting washing and separating, and culturing obtained cells to obtain first endometrial stem cells; washing endometria on the sieve, then adding a red blood cell lysate, conducting separation on lysed endometrial, then conducting shearing, digestion and separation, culturing obtained cells to obtain second endometrial stem cells; and combining the first endometrial stem cells and the second endometrial stem cells to obtain endometrial stem cells. According to the endometrial stem cell isolation method provided by the invention, menstrual blood-derived endometrial stem cells and exfoliated endometrium-derived cells areseparated, more endometrial stem cells are effectively obtained, and the endometrial stem cells meet a clinical use standard through flow cytometry detection.

Description

technical field [0001] The invention relates to the field of stem cell separation, in particular to a method for isolating endometrial stem cells. Background technique [0002] The endometrium can be divided into the basal layer and the functional layer physiologically. The basal layer includes the lower third layer of the endometrium, and its function is the place where the endometrium first develops. The underlying cells quickly regenerate the functional layer according to a specific trajectory to repair the damaged endometrium due to the shedding of the functional layer. The functional layer of the endometrium is controlled and regulated by changes in hormones in the receptors, and has certain regular changes in proliferation, differentiation and shedding. It is the place where the embryo is implanted, including the upper two-thirds of the endometrium, including the surrounding gland Loose vascularized stroma around the body and glands of pseudostratified columnar epithe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 刘年
Owner BEIJING TAIDONG BIOTECH CO LTD
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