Method for isolating uterine membrane mesenchymal stem cells by dissolving menstrual red blood cells with tea saponin

A technology for red blood cells and stem cells, applied in the biological field, can solve the problems of cell damage, affecting the number of cell cultures, etc., and achieve the effects of good cell activity, simple and controllable components, and convenient operation.

Inactive Publication Date: 2019-01-11
浙江卫未生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The Chinese patent with the patent application number 201711371272.2 discloses a method for isolating endometrial mesenchymal stem cells from menstrual blood. D...

Method used

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  • Method for isolating uterine membrane mesenchymal stem cells by dissolving menstrual red blood cells with tea saponin
  • Method for isolating uterine membrane mesenchymal stem cells by dissolving menstrual red blood cells with tea saponin
  • Method for isolating uterine membrane mesenchymal stem cells by dissolving menstrual red blood cells with tea saponin

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Embodiment 1

[0027] A method for dissolving menstrual blood red blood cells with tea saponin to separate uterine mesenchymal stem cells, comprising the steps of:

[0028] (1) Reagent preparation: prepare a tea saponin solution, the tea saponin solution is a PBS solution with a tea saponin concentration of 750 mg / L, sterilized by filtration; wherein the tea saponin powder is commercially available.

[0029] (2) Obtain cell pellet: filter 20ml of menstrual blood sample, centrifuge at 300g for 3 minutes to remove the upper layer of plasma, and obtain cell pellet A; add 45ml of tea saponin solution to 5ml of cell pellet A, mix well, let stand for 0.5-2.5min, After mixing evenly, centrifuge at 300 g for 5 min to remove the supernatant and obtain cell pellet B.

[0030] The menstrual blood sample is menstrual blood on the second or third day of the female menstrual cycle, and an equal volume of buffer solution containing at least one antibacterial substance is added, and the antibacterial substa...

Embodiment 2

[0033] A method for dissolving menstrual blood red blood cells with tea saponin to separate uterine mesenchymal stem cells, comprising the steps of:

[0034] (1) Reagent preparation: prepare a tea saponin solution, the tea saponin solution is a PBS solution with a tea saponin concentration of 375 mg / L, sterilized by filtration; wherein the tea saponin powder is commercially available.

[0035] (2) Obtain cell pellet: filter 20ml of menstrual blood sample, centrifuge at 300g for 3 minutes to remove the upper layer of plasma, and obtain cell pellet A; add 45ml of tea saponin solution to 5ml of cell pellet A, mix well, let stand for 0.5-2.5min, After mixing evenly, centrifuge at 300 g for 5 min to remove the supernatant and obtain cell pellet B.

[0036] The menstrual blood sample is menstrual blood on the second or third day of the female menstrual cycle, and an equal volume of buffer solution containing at least one antibacterial substance is added, and the antibacterial substa...

Embodiment 3

[0039] A method for dissolving menstrual blood red blood cells with tea saponin to separate uterine mesenchymal stem cells, comprising the steps of:

[0040](1) Reagent preparation: prepare a tea saponin solution, which is a PBS solution with a tea saponin concentration of 1500 mg / L, sterilized by filtration; wherein the tea saponin powder is commercially available.

[0041] (2) Obtain cell pellet: filter 20ml of menstrual blood sample, centrifuge at 300g for 3 minutes to remove the upper layer of plasma, and obtain cell pellet A; add 45ml of tea saponin solution to 5ml of cell pellet A, mix well, let stand for 0.5-2.5min, After mixing evenly, centrifuge at 300 g for 5 min to remove the supernatant and obtain cell pellet B.

[0042] The menstrual blood sample is menstrual blood on the second or third day of the female menstrual cycle, and an equal volume of buffer solution containing at least one antibacterial substance is added, and the antibacterial substance is cefotaxime s...

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Abstract

The invention relates to a method for separating uterine membrane mesenchymal stem cells by dissolving menstrual red blood cells with tea saponin, comprising the following steps: (1) preparing reagent: preparing tea saponin solution; 2, obtaining cell precipitation: filtering menstrual blood sample, centrifuging to remove upper plasma to obtain cell precipitation A; adding a tea saponin solution into the cell precipitation A, fully and uniformly mixing the tea saponin solution, removing the supernatant by centrifugation after standing, and obtaining the cell precipitation B; and (3) cell culture: suspending cell precipitate B in a cell culture medium in an incubator for adherent culture, and collecting the third passage cells to be mesenchymal stem cells. As the method adopts tea saponin,the uterine membrane tissue other than the mononuclear obtained by the traditional method can be obtain without damaging the target cells, and the uterine membrane mesenchymal stem cells can be obtainby inducing the mononuclear cells and the uterine membrane tissue from the menstrual blood rapidly and efficiently, and the number is several times of the of the traditional white membrane layer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for dissolving menstrual red blood cells with tea saponin and isolating uterine mesenchymal stem cells. Background technique [0002] Stem cells are a type of cells with the ability of self-renewal, proliferation and differentiation, and have great application value in cell and tissue repair, as well as as a carrier of gene therapy. Mesenchym stem cells (MesenchymAlstemcells, MSCs) have a wide range of sources, strong plasticity, and have many advantages such as easy isolation and culture, multiple factor secretion functions, and immune regulation functions. They are currently a research hotspot in the field of stem cells. Many studies have shown that MSCs have good application prospects in the repair of tissue damage and the treatment of various diseases, including participating in the repair of tissue defects such as bone, cartilage and tendon, improving cardiac function i...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2509/00
Inventor 孙巍群陈锦阳刘军权
Owner 浙江卫未生物医药科技有限公司
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