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Method for culturing menstrual blood mesenchymal stem cells

A mesenchymal stem cell and stem cell technology, applied in the field of preparation of menstrual blood mesenchymal stem cells, can solve problems such as damage, achieve the effect of maintaining cell shape, maintaining telomerase expression, and increasing production

Active Publication Date: 2020-08-25
SHENZHEN BEIKE BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But obtaining these cells directly is a very damaging process

Method used

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  • Method for culturing menstrual blood mesenchymal stem cells
  • Method for culturing menstrual blood mesenchymal stem cells
  • Method for culturing menstrual blood mesenchymal stem cells

Examples

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Embodiment 1

[0041] Example 1 Preparation of decidual mesenchymal stem cells

[0042] The method for preparing menstrual blood mesenchymal stem cells in this embodiment comprises the following steps:

[0043] Use a women-specific menstrual cup to collect women's menstrual blood on the second or third day, and collect 10-12ml of menstrual blood each time. Immediately pour the collected menstrual blood into a collection bottle filled with preservation solution, store it at 4°C, and send it to the laboratory for processing as soon as possible. In the laboratory, the samples were subjected to differential centrifugation and washed to remove bacteria as much as possible, and then hydroxyethyl starch (HES) was added to obtain mononuclear cells in menstrual blood for culturing mesenchymal stem cells.

[0044] The primary culture method is:

[0045](1) Use a menstrual cup to collect women's menstrual blood for a day, collect 2-3 times, and collect 3-15ml of menstrual blood each time. Immediatel...

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Abstract

The invention provides a method for preparing menstrual blood mesenchymal stem cells, which comprises collecting women's menstrual blood as a raw material, aseptically treating menstrual blood; separating stem cells in menstrual blood; cultivating menstrual blood mesenchymal stem cells; freezing and storing menstrual blood mesenchymal stem cells; Recovery of mesenchymal stem cells. The method of the present invention has the following technical features: collect women's menstrual blood, and preserve the liquid in the preservation liquid; prevent bacterial contamination, reduce the probability of contamination from the source of collection, and rinse the collection cup with sterile water for multiple times, effectively reducing the probability of contamination; The differential centrifugation method removes bacteria in menstrual blood as much as possible; repeated separation of samples by hydroxyethyl starch (HES) can obtain the largest number of mesenchymal stem cells in menstrual blood; use serum-free medium for culture to reduce the use of animal-derived components, The cell performance is stable, and it can maintain the long-term culture process of menstrual blood mesenchymal stem cells in vitro, maintain cell shape, proliferation ability, expression of MSC surface markers, differentiation ability, etc. The method used is simple and easy to operate, and can obtain the maximum amount of required stem cells and successfully cultivate them.

Description

technical field [0001] The invention relates to a method for preparing menstrual blood mesenchymal stem cells, belonging to the technical field of stem cells Background technique [0002] Embryonic stem cells (ESCs) are pluripotent and can differentiate into multiple cell types of all three germ layer cell layers. However, the teratogenic potential of embryonic stem cells and their limitations in large-scale clinical applications severely limit their use in scientific research. However, with the increasing number of patients suffering from myriad conditions, we are urgently looking for other sources of stem cells with clinical value. Many other sources of stem cells can be used in early clinical trials, such as heart disease, spinal cord injury, bone and cartilage repair, etc. Stromal stem cell fragments from many tissues and organs exhibit multidifferentiation potential in vitro, as they can differentiate into cell lines such as neurogenic and cardiogenic, osteogenic, cho...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775
Inventor 曾桂芳李陶杨威仁何毅欣胡祥刘沐芸
Owner SHENZHEN BEIKE BIOTECH
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