Multifunctional menstrual blood stem cell culture method

Abstract

The invention relates to a multifunctional menstrual blood stem cell culture method which comprises the following steps: menstrual blood and plasma acquisition, menstrual blood bactericidal cleaning, menstrual blood filtration, autoplasma preparation, single karyocyte separation, menstrual blood stem cell culture, menstrual blood menstruation purification and amplification, and menstrual blood stem cell cryopreservation. The method overcomes the defects caused by the required use of high-concentration fetal calf serum for culture in the prior art; and by using the own plasma of the customer to culture the cells, the customer can feel safe and acceptable, thereby implementing the safe, stable and quick high-quality culture system for amplifying the mesenchymal stem cells in deed.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a multifunctional method for culturing menstrual blood stem cells, in particular to a method for extracting and culturing menstrual blood stem cells from menstrual blood. Background technique

[0002] It is well known that the endometrial cell line has a strong self-renewal ability, and there will be a large growth of tissue and blood vessels in each menstrual cycle. This growth process will stop with the onset of menstruation and then form menstrual blood. Menstrual blood is a mixture of a woman's blood and some shed uterine lining, cervical mucus, and vaginal discharge. In recent years, research groups in the United States and Japan have successfully isolated and cultured stem cells with multi-directional differentiation functions using women's menstrual blood, and named the cells as endometrial menstrual blood-derived mesenchymal stem cells, referred to as menstrual bl...

Examples

Embodiment 1

[0030] A multifunctional method for culturing menstrual blood stem cells, the steps are as follows:

[0031] Step 1: The collection stage of menstrual blood and plasma. This stage is specifically: use a menstrual blood collector to collect menstrual blood. The selected menstrual blood collector should be non-invasive to the female vulva, easy to use, not easy to be polluted, and low in cost. Stem cells are extracted for the next step Provides high-quality assurance, and quickly transfers menstrual blood to a sterile centrifuge tube filled with preservation solution, and stores it at a temperature of 4°C. Take 20ml of venous blood on an empty stomach and put it into an anticoagulant container. After being stored at ℃ for 72 hours, it is used for the preparation of autologous plasma;

[0032] Step 2: Sterilization and cleaning of menstrual blood. This stage is specifically: transport the menstrual blood soaked in the menstrual blood preservation solution to the laboratory within...

Embodiment 2

[0051] A multifunctional method for culturing menstrual blood stem cells, the steps are as follows:

[0052] Step 1: The collection stage of menstrual blood and plasma. This stage is specifically: collect menstrual blood with a menstrual blood collector, and quickly transfer the menstrual blood to a sterile centrifuge tube filled with preservation solution, store it at a temperature of 4°C, and fast Take 25ml of venous blood and place it in an anticoagulant container, store it at 4°C for 72 hours, and use it for the preparation of autologous plasma;

[0053] Step 2: The stage of sterilization and cleaning of menstrual blood. This stage is specifically: transport the menstrual blood soaked in the menstrual blood preservation solution to the laboratory within 48 hours, fully mix the menstrual blood and the preservation solution, and centrifuge at 900g / 13min for the first time, remove For the supernatant, add the cleaning solution again, mix well, centrifuge again to 500g / 13min, ...

Embodiment 3

[0072] A multifunctional method for culturing menstrual blood stem cells, the steps are as follows:

[0073] Step 1: The collection stage of menstrual blood and plasma. This stage is specifically: collect menstrual blood with a menstrual blood collector, and quickly transfer the menstrual blood to a sterile centrifuge tube filled with preservation solution, store it at a temperature of 4°C, and fast Draw 30ml of venous blood into an anticoagulant container, store it at 4°C for 72 hours, and use it for the preparation of autologous plasma;

[0074] Step 2: The stage of sterilization and cleaning of menstrual blood. This stage is specifically: transport the menstrual blood soaked in the menstrual blood preservation solution to the laboratory within 48 hours, fully mix the menstrual blood and the preservation solution, and centrifuge at 1000g / 15min for the first time, remove For the supernatant, add the cleaning solution again, mix well, centrifuge again to 700g / 15min, and remove...

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a multifunctional method for culturing menstrual blood stem cells, in particular to a method for extracting and culturing menstrual blood stem cells from menstrual blood. Background technique

[0002] It is well known that the endometrial cell line has a strong self-renewal ability, and there will be a large growth of tissue and blood vessels in each menstrual cycle. This growth process will stop with the onset of menstruation and then form menstrual blood. Menstrual blood is a mixture of a woman's blood and some shed uterine lining, cervical mucus, and vaginal discharge. In recent years, research groups in the United States and Japan have successfully isolated and cultured stem cells with multi-directional differentiation functions using women's menstrual blood, and named the cells as endometrial menstrual blood-derived mesenchymal stem cells, referred to as menstrual bl...

Examples

Embodiment 1

[0030] A multifunctional method for culturing menstrual blood stem cells, the steps are as follows:

[0031] Step 1: The collection stage of menstrual blood and plasma. This stage is specifically: use a menstrual blood collector to collect menstrual blood. The selected menstrual blood collector should be non-invasive to the female vulva, easy to use, not easy to be polluted, and low in cost. Stem cells are extracted for the next step Provides high-quality assurance, and quickly transfers menstrual blood to a sterile centrifuge tube filled with preservation solution, and stores it at a temperature of 4°C. Take 20ml of venous blood on an empty stomach and put it into an anticoagulant container. After being stored at ℃ for 72 hours, it is used for the preparation of autologous plasma;

[0032] Step 2: Sterilization and cleaning of menstrual blood. This stage is specifically: transport the menstrual blood soaked in the menstrual blood preservation solution to the laboratory within...

Embodiment 2

[0051] A multifunctional method for culturing menstrual blood stem cells, the steps are as follows:

[0052] Step 1: The collection stage of menstrual blood and plasma. This stage is specifically: collect menstrual blood with a menstrual blood collector, and quickly transfer the menstrual blood to a sterile centrifuge tube filled with preservation solution, store it at a temperature of 4°C, and fast Take 25ml of venous blood and place it in an anticoagulant container, store it at 4°C for 72 hours, and use it for the preparation of autologous plasma;

[0053] Step 2: The stage of sterilization and cleaning of menstrual blood. This stage is specifically: transport the menstrual blood soaked in the menstrual blood preservation solution to the laboratory within 48 hours, fully mix the menstrual blood and the preservation solution, and centrifuge at 900g / 13min for the first time, remove For the supernatant, add the cleaning solution again, mix well, centrifuge again to 500g / 13min, ...

Embodiment 3

[0072] A multifunctional method for culturing menstrual blood stem cells, the steps are as follows:

[0073] Step 1: The collection stage of menstrual blood and plasma. This stage is specifically: collect menstrual blood with a menstrual blood collector, and quickly transfer the menstrual blood to a sterile centrifuge tube filled with preservation solution, store it at a temperature of 4°C, and fast Draw 30ml of venous blood into an anticoagulant container, store it at 4°C for 72 hours, and use it for the preparation of autologous plasma;

[0074] Step 2: The stage of sterilization and cleaning of menstrual blood. This stage is specifically: transport the menstrual blood soaked in the menstrual blood preservation solution to the laboratory within 48 hours, fully mix the menstrual blood and the preservation solution, and centrifuge at 1000g / 15min for the first time, remove For the supernatant, add the cleaning solution again, mix well, centrifuge again to 700g / 15min, and remove...

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