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Culturing method of mesenchymal stem cells of menstrual blood

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

Active Publication Date: 2015-04-29
SHENZHEN BEIKE BIOTECH
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Problems solved by technology

But obtaining these cells directly is a very damaging process

Method used

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  • Culturing method of mesenchymal stem cells of menstrual blood
  • Culturing method of mesenchymal stem cells of menstrual blood
  • Culturing method of mesenchymal stem cells of menstrual blood

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. Imme...

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Abstract

The invention provides a method for preparing mesenchymal stem cells of menstrual blood. The method comprises steps as follows: female menstrual blood is collected and taken as a raw material and is subjected to sterile processing; stem cells in the menstrual blood are separated; the mesenchymal stem cells of the menstrual blood are cultured; the mesenchymal stem cells of the menstrual blood are cryopreserved; and the mesenchymal stem cells of the menstrual blood are recovered. The method has the technical characteristics as follows: the female menstrual blood is collected and stored in a preservative fluid; by means of a bacterial pollution prevention method, the pollution probability is reduced from a collection source, a collecting cup is repeatedly washed by sterile water, and the pollution probability is effectively reduced; with the adoption of a differential centrifugation method, bacteria in the menstrual blood are removed as far as possible; a sample is repeatedly separated by HES (hydroxyethyl starch), and the maximum quantity of mesenchymal stem cells of the menstrual blood can be obtained; a serum-free medium is utilized for culturing, components of animal origin are reduced, the cell performance is stable, the in-vitro long-term culturing process of the mesenchymal stem cells of the menstrual blood can be kept, and cellular morphology, multiplication capacity, MSC (mesenchymal stem cell) surface marker expression, differentiation capacity and the like are maintained. The method is simple, practical and convenient to operate, the maximum quantity of required stem cells can be obtained, and the stem cells are successfully cultured.

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 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, chondrog...

Claims

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Application Information

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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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