CN113755433A - Synovial mesenchymal stem cell suspension and preparation method thereof - Google Patents
Synovial mesenchymal stem cell suspension and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a synovial membrane mesenchymal stem cell suspension and a preparation method thereof, belonging to the technical field of medical manufacturing. It comprises the following steps: (1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell; (2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture; (3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell. The preparation method of the synovial mesenchymal stem cell suspension is simple in process and easy to manufacture, and is beneficial to improving the suspension stability of the synovial mesenchymal stem cells.
Description
Technical Field
The invention belongs to the technical field of medical manufacturing, and particularly relates to a synovial mesenchymal stem cell suspension and a preparation method thereof.
Background
Mesenchymal Stem Cells (MSCs) refer to clonal cells derived from mesenchymal tissue with high proliferative and multipotent differentiation potential. A large number of researches prove that the MSCs have the directional differentiation capacity to adipocytes, osteoblasts, chondrocytes, myocytes and the like, and increasingly show wide application prospects in the fields of tissue engineering, regenerative medicine and the like.
And is considered to be the most ideal seed cell for repairing cartilage damage due to the excellent chondrogenic differentiation potential. However, the related studies of mesenchymal stem cells are relatively rare and the biological characteristics and possible effects are poorly understood. Currently, synovial mesenchymal stem cells of mice can be used for simulation experiment research. For example, the Chinese patent, application number: cn201810155580.x, publication No.: CN108456655A discloses a mesenchymal stem cell suspension, a preparation method and an application thereof, and the technical proposal is as follows: obtaining a primary mesenchymal stem cell; culturing and amplifying the primary mesenchymal stem cell: after 5-10 days of culture of the primary mesenchymal stem cells, the primary mesenchymal stem cells are replaced by serum-free DMEM nutrient solution until 65-75% of the cells are fused, culture supernatant is removed, first digestive fluid is added, the cells are digested for 0.5-2min, the cells are added into the removed culture supernatant after being contracted and then are neutralized for centrifugal separation, complete culture medium is added again for heavy suspension, inoculation is carried out according to the 1 st transmission rate of 1-1.5, after 4-5 days of culture, the cells are fused at the ratio of 1: subculturing at a ratio of 6-8; and preparing a mesenchymal stem cell suspension: and (3) taking P4 generation cells in the culture and amplification of the primary mesenchymal stem cell to culture and amplify for 48-96 hours, after the cells are fused for no more than 80%, digesting the cells by using a third digestion solution, washing, suspending by using 5% human albumin, and adding physiological saline to obtain the required mesenchymal stem cell suspension. However, the patent does not modify DMEM, resulting in short shelf life and limited cell viability.
Disclosure of Invention
1. Problems to be solved
Aiming at the problems in the prior art, the invention provides the synovial membrane mesenchymal stem cell suspension and the preparation method thereof, which have the advantages of simple process and easy manufacture and are beneficial to improving the suspension stability of the synovial membrane mesenchymal stem cells.
2. Technical scheme
In order to solve the above problems, the present invention adopts the following technical solutions.
A preparation method of a synovial mesenchymal stem cell suspension comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
The synovial mesenchymal stem cell suspension is prepared by the preparation method of the synovial mesenchymal stem cell suspension.
3. Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
the improved DMEM culture medium is innovatively introduced, the new additive zinc acetate plays a critical role, and the hyaluronic acid also plays a certain promoting role.
Drawings
FIG. 1 is a graph showing the results of laser confocal in example 1;
FIG. 2 is a graph showing the results of laser confocal in comparative example 1;
FIG. 3 is a graph showing the results of laser confocal in comparative example 2;
FIG. 4 is a graph showing the results of laser confocal in comparative example 3;
FIG. 5 is a graph showing the results of laser confocal in comparative example 4;
FIG. 6 is a graph showing the results of laser confocal in comparative example 5.
Detailed Description
The invention is further described with reference to specific examples.
It needs to be reminded that the synovial membrane mesenchymal stem cells researched by the application are mouse synovial membrane mesenchymal stem cells, are mainly used for researching the suspension stability of the mouse synovial membrane mesenchymal stem cells, and are combined with instruments such as a live-dead cell staining kit to detect the activity of the mouse synovial membrane mesenchymal stem cells.
Example 1
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 1
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 2
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
trypsin 1 mg.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 3
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
zinc acetate 0.2 mg.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 4
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
and recombining into fiber growth factor FGF-9100 ng.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Comparative example 5
The preparation method of the synovial mesenchymal stem cell suspension of the embodiment comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing into a DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the condition parameters of the second culture in the step (2) are as follows:
the cell fusion phenomenon occurs at 70% in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃, a DMEM medium and a 7d culture.
In the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃;
in the above-mentioned method for preparing a synovial mesenchymal stem cell suspension,
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
In the preparation method of the synovial mesenchymal stem cell suspension, the temperature for treatment in the step (3) is 10 ℃.
In the preparation method of the synovial mesenchymal stem cell suspension, the preservation temperature after the treatment in the step (3) is 4 ℃.
Example 6
The suspension prepared in example 1 and the suspensions prepared in comparative examples 1 to 5 were selected and tested as follows:
the suspension prepared above was stored at 4 ℃ for one month, and then the identification of stem cells in the suspension was observed using a live-dead cell staining kit (purchased from Eimei technologies, Inc., Wuhan) in combination with a laser confocal instrument.
As shown in fig. 1, the number of living cells is large, the number of dead cells is small, the morphology is normal, and the cell density is normal;
as shown in FIG. 2, the number of living cells is less than that of FIG. 1, the number of dead cells is more than that of FIG. 1, the morphology of part of the cells is distorted, the number of dead cells is more than that of living cells as a whole, and the cell density is less than that of FIG. 1;
as shown in fig. 3, the number of living cells is less than that of fig. 2, the number of dead cells is more than that of fig. 2, the number of dead cells is much more than that of living cells as a whole, the morphology of part of the cells is distorted, and the cell density is less than that of fig. 1;
as shown in fig. 4, the number of living cells was the smallest, the number of dead cells was greater than that of fig. 3, and the number of dead cells was 5 times as large as that of living cells as a whole, and the cell density was the smallest;
as shown in FIG. 5, the number of live cells is less than that of FIG. 3 and more than that of FIG. 4, the number of dead cells is 2 times that of dead cells as a whole, and the cell density is less than that of FIG. 4 and less than that of FIG. 3;
as shown in fig. 6, the number of live cells was between fig. 4 and fig. 5, the number of dead cells was equal to the number of live cells as a whole, and the cell density was between fig. 4 and fig. 5.
While the invention has been described in further detail in connection with specific embodiments thereof, it will be understood that the invention is not limited thereto, and that various other modifications and substitutions may be made by those skilled in the art without departing from the spirit of the invention, which should be considered to be within the scope of the invention as defined by the appended claims.
Claims (10)
1. A method for preparing a suspension of synovial mesenchymal stem cells, which is characterized by comprising the following steps:
the method comprises the following steps:
(1) pretreatment: taking a commercialized packaging cell of the synovial mesenchymal stem cell;
(2) culturing: performing amplification culture on the synovial membrane mesenchymal stem cells in the step (1), and obtaining cells after subculture;
(3) preparation of the suspension: and (3) transferring the cells subjected to subculture in the step (2) into a treatment solution for treatment to obtain the cell.
2. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the synovial membrane mesenchymal stem cells in the step (1) are mouse synovial membrane mesenchymal stem cells, and the stock number of the synovial membrane mesenchymal stem cells is as follows: CP-M236, and was purchased from Wuhan Ponno Life technologies, Inc.
3. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the amplification culture method in the step (2) is as follows:
taking the synovial membrane mesenchymal stem cells in the step (1) as primary cells, carrying out primary culture, then changing to an improved DMEM culture medium for secondary culture, then adding trypsin with the mass fraction of 0.50% for digestion treatment, wherein the treatment temperature is 37 ℃, then carrying out centrifugal treatment at 4 ℃, and taking precipitated cells.
4. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 3, wherein:
the condition parameters of the first culture in the step (2) are as follows:
culturing in a carbon dioxide incubator with the volume fraction of 5% at 37 ℃ for 5 d;
the basic culture medium is HG-DMEM culture medium, and fetal bovine serum, penicillin and streptomycin are added into the culture medium, wherein the addition amount of the fetal bovine serum is 10% of the volume of the HG-DMEM culture medium, the addition amount of the penicillin is 100U/mL, and the addition amount of the streptomycin is 100U/mL.
5. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the condition parameters of the second culture in the step (2) are as follows:
the carbon dioxide incubator with 5% volume fraction, the modified DMEM medium and the 7d culture are used for carrying out 70% fusion phenomenon on the cells at 37 ℃;
the components of the modified DMEM medium are as follows:
the DMEM culture medium is 100mL,
reconstituting the fiber growth factor FGF-4100 ng,
reconstituting the fiber growth factor FGF-9100 ng,
zinc acetate 0.2 mg.
6. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the subculture method in step (2) is as follows:
the cells after expanded culture were transferred to subculture medium at a ratio of 1: 3, subculturing, and replacing the subculture medium every 3 d; wherein the subculture medium is a DMEM medium; the condition parameters of subculture are as follows:
a carbon dioxide incubator with the volume fraction of 5 percent at 37 ℃.
7. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the treating fluid in the step (3) comprises the following components:
the volume of the physiological saline is 100mL,
1mg of trypsin is added to the mixture,
hyaluronic acid 0.3 mL.
8. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the temperature for the treatment in step (3) was 10 ℃.
9. The method for preparing a suspension of synovial mesenchymal stem cells according to claim 1, wherein:
the preservation temperature after the treatment in step (3) was 4 ℃.
10. A suspension of synovial mesenchymal stem cells, wherein:
the synovial mesenchymal stem cell suspension of claim 1.
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115418348A (en) * | 2022-08-06 | 2022-12-02 | 北京永华燕芸生物科技有限公司 | Method for extracting and culturing synovial effusion mesenchymal stem cells |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102978156A (en) * | 2012-11-13 | 2013-03-20 | 湖州市中心医院 | Expansion in vitro purification culture method of mesenchymal stem cells and culture medium |
US20150086514A1 (en) * | 2012-11-14 | 2015-03-26 | Zaimei JIA | Mesenchymal stem cell injection and preparation method thereof, and application thereof in preparing diabetes drug |
CN105524878A (en) * | 2016-01-21 | 2016-04-27 | 深圳市第二人民医院 | Culture method for separating and inducting hUC-MSCs into cartilage cells |
CN108456655A (en) * | 2018-02-23 | 2018-08-28 | 深圳至博生物科技有限公司 | Mescenchymal stem cell suspension and the preparation method and application thereof |
-
2021
- 2021-08-09 CN CN202110907400.0A patent/CN113755433A/en active Pending
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102978156A (en) * | 2012-11-13 | 2013-03-20 | 湖州市中心医院 | Expansion in vitro purification culture method of mesenchymal stem cells and culture medium |
US20150086514A1 (en) * | 2012-11-14 | 2015-03-26 | Zaimei JIA | Mesenchymal stem cell injection and preparation method thereof, and application thereof in preparing diabetes drug |
CN105524878A (en) * | 2016-01-21 | 2016-04-27 | 深圳市第二人民医院 | Culture method for separating and inducting hUC-MSCs into cartilage cells |
CN108456655A (en) * | 2018-02-23 | 2018-08-28 | 深圳至博生物科技有限公司 | Mescenchymal stem cell suspension and the preparation method and application thereof |
Non-Patent Citations (9)
Title |
---|
HIDEYA YOSHIMURA等: "《Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle》", 《CELL TISSUE RES.》 * |
JI HYUN KIM等: "《Enhanced proliferation and chondrogenic differentiation of human synovium-derived stem cells expanded with basic fibroblast growth factor》", 《TISSUE ENG PART A.》 * |
MI-YOUNG MOON等: "《Zinc Promotes Adipose-Derived Mesenchymal Stem Cell Proliferation and Differentiation towards a Neuronal Fate》", 《STEM CELLS INTERNATIONAL》 * |
SMITHA MATHEWS等: "《Glycosaminoglycans enhance osteoblast differentiation of bone marrow derived human mesenchymal stem cells》", 《JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE》 * |
庞雪芬: "《透明质酸对脂肪干细胞增殖和分化的影响》", 《中山大学硕士学位论文》 * |
张正政等: "《滑膜间充质干细胞分离培养、免疫表型鉴定及在混合淋巴反应体系中的抑制效应》", 《中国组织工程研究》 * |
张正政等: "滑膜间充质干细胞分离培养、免疫表型鉴定及在混合淋巴反应体系中的抑制效应", 《中国组织工程研究》 * |
符培亮等: "滑膜间充质干细胞分离纯化后的生物学特性", 《中国组织工程研究》 * |
龚忠诚等: "滑膜间充质干细胞软骨分化能力的实验研究", 《新疆医科大学学报》 * |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN115418348A (en) * | 2022-08-06 | 2022-12-02 | 北京永华燕芸生物科技有限公司 | Method for extracting and culturing synovial effusion mesenchymal stem cells |
CN115418348B (en) * | 2022-08-06 | 2024-05-31 | 北京永华燕芸生物科技有限公司 | Extraction and culture method of synovial effusion mesenchymal stem cells |
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