Preparation method and application method of umbilical cord blood-derived mesenchymal mononuclear cells for treating urinary incontinence

Abstract

The invention discloses a preparation method and an application method of umbilical cord blood-derived mesenchymal mononuclear cells for treating urinary incontinence, and particularly relates to the field of medical equipment.The preparation method comprises the following steps that magnetic powder is selected and mixed with drugs added for preparing the mononuclear cells; preparing mixed medicine powder; s2, putting the mixed medicine powder in the step S2 into a centrifugal tube, and preserving under a vacuum condition to obtain a preserved sample; and putting the preserved sample into a centrifugal tube, and preserving under a vacuum condition to obtain a round-cake-shaped umbilical cord monocyte sample. The low-cost Y-Gd-based alloy is used for modifying the magnet mononuclear cells, the alternating current impedance spectrum radius of the prepared umbilical cord mononuclear cells is increased, the corrosion degree in a corrosive environment is reduced, the corrosion resistance of the magnet is enhanced, and the problem that the magnet is difficult to use for a long time in a corrosive medium and a humid and hot environment is effectively solved.

Description

Technical Field

[0001] The present invention relates to the technical field of medical equipment, and more specifically, to a preparation method and application method of mesenchymal mononuclear cells derived from umbilical cord blood for treating urinary incontinence. Background Art

[0002] Stress urinary incontinence (SUI) is a common disease in middle-aged and elderly women, with an incidence rate of 10-23% in Europe and the United States and about 18.9% in China. The causes of female stress urinary incontinence are: 1. After the age of 40, women's ovarian function begins to decline, estrogen decreases, and epithelial cells begin to atrophy; 2. Muscle damage during pregnancy and childbirth; 3. Congenital developmental factors; Symptoms include involuntary leakage of urine when abdominal pressure increases (coughing, sneezing, laughing, carrying weight, etc.), which brings great trouble to the daily life of middle-aged and elderly women and leads to a significant decline in their quality of life.

[0003] At present, the treatments for female stress urinary incontinence include physical exercise, drug therapy and surgical treatment. When the symptoms are mild, physical exercise has a certain effect, and drug therapy is controversial due to its large side effects and poor efficacy. Surgical treatment should be the most effective method at present. Surgical treatment includes three types: anterior vaginal wall repair, retropubic urethral bladder suspension, and sling surgery. Because it is a traumatic surgery, improper operation will cause side effects such as tissue damage, postoperative pain and discomfort, and urinary retention. Therefore, there is an urgent need to prepare a method and application method of umbilical cord blood-derived mesenchymal monocytes for the treatment of urinary incontinence. Summary of the invention

[0004] The present invention provides a preparation method and application method of umbilical cord blood-derived mesenchymal mononuclear cells for treating urinary incontinence, and aims to solve the problem that existing umbilical cord mononuclear cells have poor corrosion resistance and are not conducive to long-term use in corrosive media and hot and humid environments.

[0005] To achieve the above object, the present invention provides the following technical solution: a preparation method and application method of umbilical cord blood-derived mesenchymal stem cells for treating urinary incontinence, comprising the following steps:

[0006] S1: Selecting Ce-rich main phase MQP magnetic powder and Nd-rich main phase MQU-F magnetic powder in the same proportion, and preparing drugs added to mononuclear cells;

[0007] S2: mixing the same proportion of Ce-rich main phase MQP magnetic powder and Nd-rich main phase MQU-F magnetic powder with 0-4wt% of the total drug to obtain a mixed drug powder;

[0008] S3: placing the mixed drug powder in step S2 into a centrifuge tube and storing it under vacuum conditions to obtain a stored sample;

[0009] S4: placing the preserved sample into a centrifuge tube and preserving it under vacuum conditions to obtain a round cake-shaped umbilical cord mononuclear cell sample;

[0010] The molecular formula of the Ce-rich main phase MQP magnetic powder in step S1 is Nd 29.8 Pr 0.2 Ga 0.46 Fe bal. Co4B 0.94 ; The molecular formula of Nd-rich main phase MQU-F magnetic powder is Ce 23.6 NdP 1.9 Fe 69.5 B; The molecular formula of the drug added to the monocytes is Gd 60 Y 10 Cu 15 Al 15 .

[0011] In a preferred embodiment, the method for preparing the drug added to monocytes in step S1 further comprises the following steps:

[0012] S11: vacuum melting Y, Gd, Cu and Al elements according to the molecular ratio of the alloy material to obtain an alloy ingot;

[0013] S12: subjecting the alloy ingot in step S11 to vacuum quenching and strip spinning in a vacuum strip spinning machine to obtain an alloy strip;

[0014] S13: Grind the alloy strip in step S12 into powder to obtain alloy medicine powder.

[0015] In a preferred embodiment, the method for preparing the mixed drug powder in step S2 further comprises the following steps:

[0016] S21: first weigh 50 parts by weight of Ce-rich main phase MQP magnetic powder and 50 parts by weight of Nd-rich main phase MQU-F magnetic powder;

[0017] S22: Then, the drug with an addition amount of 1wt%, 2wt%, 3wt% or 4wt% is uniformly mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder to prepare mixed drug powders containing different contents of alloy substances.

[0018] In a preferred embodiment, in step S3, the method for preserving the sample further comprises the following steps:

[0019] S31: first pour the mixed drug powder into a centrifuge tube with a diameter of φ20 for storage;

[0020] S32: The mixed drug powder is then placed in the center of the furnace of the centrifuge tube, and after vacuuming, the heating and pressurization procedures are started simultaneously.

[0021] In a preferred embodiment, the method for preserving and molding the umbilical cord mononuclear cell sample in step S4 further comprises the following steps:

[0022] S41: first take out the dense precursor sample in step S3 and put it into a φ36 centrifuge tube;

[0023] S42: Then, the mixed drug powder is placed in the center of the furnace of the centrifuge tube, and after vacuuming, heating is started first.

[0024] In a preferred embodiment, in step S4, the deformation amount of the Cerium Iron Boron magnet sample is 70%, and the outer circle size is φ36 and the height is 8mm in a round cake-shaped structure.

[0025] In a preferred embodiment, the temperature parameters of the storage process in step S32 are set as follows: heating to 300°C at an output power of 30W, then heating to 550°C at an output power of 50W, keeping warm for 10 minutes, then continuing to heat to 680°C at this power, and finally keeping warm for 30 minutes.

[0026] In a preferred embodiment, the process pressure parameter settings saved in step S32 are: pressurizing to 60MPa, followed by maintaining the pressure for 30 minutes. The process parameters saved in step S42 are set as: heating to 300°C at an output power of 30W, then heating to 650°C at an output power of 50W, maintaining the temperature for 10 minutes, and then continuing to heat to 750°C at this power, and finally maintaining the temperature for 20 minutes. When the temperature reaches 750°C, the pressure is started, and it takes 20 minutes (deformation rate is 0.001s-1) to increase the pressure to 90MPa.

[0027] A kind of umbilical cord mononuclear cell, the atomic percentage of Nd-Ce-Fe-B magnetic powder is Nd x Ce y Fe 100-x-y-z-n B z M n , where 12≤x+y≤16, 5.5≤z≤7, 0.1≤n≤5, and M is one or more of Co, Ni, Al, Cu, Ga, Mg, Zn, Nb, Zr, Ti, Mo, W, and V elements.

[0028] Technical effects and advantages of the present invention:

[0029] The present invention uses low-cost Y-Gd-based alloy to modify multi-phase (Nd, Ce)-Fe-B magnet mononuclear cells, and the prepared umbilical cord mononuclear cells have a larger AC impedance spectrum radius, a reduced degree of corrosion in a corrosive environment, and enhanced corrosion resistance of the magnet, effectively overcoming the problem that the magnet is difficult to use for a long time in a corrosive medium and a hot and humid environment. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is a schematic diagram of the temperature and pressure parameters for storage during the preparation of umbilical cord mononuclear cells of the present invention;

[0031] Figure 2 A schematic diagram of the open circuit potential of the side of the umbilical cord mononuclear cells prepared by adding different proportions of Y-Gd-based alloys in the present invention in a 3.5wt% NaCl corrosion solution;

[0032] Figure 3 The schematic diagram of the Nyquist curve and polarization curve of the AC impedance spectrum of the umbilical cord mononuclear cells prepared by adding different proportions of Y-Gd-based alloys in the present invention;

[0033] Figure 4 Schematic diagram of the AC impedance spectrum and polarization curve of the side of the umbilical cord mononuclear cells prepared in each embodiment of the present invention in a 3.5wt% NaCl solution;

[0034] Figure 5 Schematic diagram of corrosion current and current density on the side of umbilical cord mononuclear cells prepared in each embodiment of the present invention;

[0035] Figure 6 Schematic diagram of the corrosion morphology of umbilical cord mononuclear cells prepared in each embodiment of the present invention;

[0036] Figure 7 This is a schematic diagram of the corrosion morphology of the side of the umbilical cord mononuclear cell prepared without adding Y-Gd alloy in the present invention;

[0037] Figure 8 This is a schematic diagram of the side corrosion morphology of umbilical cord mononuclear cells prepared by adding 2wt% Y-Gd alloy in the present invention. DETAILED DESCRIPTION

[0038] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0039] Example 1

[0040] A preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence, comprising the following steps:

[0041] S1: Selecting Ce-rich main phase MQP magnetic powder and Nd-rich main phase MQU-F magnetic powder in the same proportion, and preparing drugs added to mononuclear cells;

[0042] S11: vacuum melting Y, Gd, Cu and Al elements according to the molecular ratio of the alloy material to obtain an alloy ingot;

[0043] S12: subjecting the alloy ingot in step S11 to vacuum quenching and strip spinning in a vacuum strip spinning machine to obtain an alloy strip;

[0044] S13: Grind the alloy strip in step S12 into powder to obtain alloy medicine powder.

[0045] S2: mixing the same proportion of Ce-rich main phase MQP magnetic powder and Nd-rich main phase MQU-F magnetic powder with 0-4wt% of the total drug to obtain a mixed drug powder;

[0046] S21: first weigh 50 parts by weight of Ce-rich main phase MQP magnetic powder and 50 parts by weight of Nd-rich main phase MQU-F magnetic powder;

[0047] S22: Then, the drug with an addition amount of 0 wt% is uniformly mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder respectively to prepare mixed drug powders containing different contents of alloy substances.

[0048] S3: placing the mixed drug powder in step S2 into a centrifuge tube and storing it under vacuum conditions to obtain a stored sample;

[0049] S31: first pour the mixed drug powder into a centrifuge tube with a diameter of φ20 for storage;

[0050] S32: The mixed drug powder is then placed in the center of the furnace of the centrifuge tube, and after vacuuming, the heating and pressurization procedures are started simultaneously.

[0051] The temperature parameters of the storage process in step S32 are set as follows: heating to 300°C at an output power of 30W, then heating to 550°C at an output power of 50W, keeping warm for 10 minutes, then continuing to heat to 680°C at this power, and finally keeping warm for 30 minutes; the pressure parameters of the storage process in step S32 are set as follows: pressurizing to 60MPa, and then keeping the pressure for 30 minutes.

[0052] S4: placing the preserved sample into a centrifuge tube and preserving it under vacuum conditions to obtain a round cake-shaped umbilical cord mononuclear cell sample;

[0053] S41: first take out the dense precursor sample in step S3 and put it into a φ36 centrifuge tube;

[0054] S42: Then, the mixed drug powder is placed in the center of the furnace of the centrifuge tube, and after vacuuming, heating is started first;

[0055] The saved process parameters in step S42 are set as follows: heating to 300°C at an output power of 30W, then heating to 650°C at an output power of 50W, keeping warm for 10 minutes, then continuing to heat to 750°C at this power, and finally keeping warm for 20 minutes. When the temperature reaches 750°C, the pressure is started, and it takes 20 minutes (deformation rate is 0.001s-1) to increase the pressure to 90MPa.

[0056] Recorded as control group 1.

[0057] After the thermal deformation, a round-shaped cerium-iron-boron magnet sample with a deformation amount of 70%, an outer circle size of φ36, and a height of 8 mm was finally obtained.

[0058] The molecular formula of the Ce-rich main phase MQP magnetic powder in step S1 is Nd 29.8 Pr 0.2 Ga 0.46 Fe bal. Co4B 0.94 ; The molecular formula of Nd-rich main phase MQU-F magnetic powder is Ce 23.6 NdP 1.9 Fe 69.5 B; The molecular formula of the drug added to the monocytes is Gd 60 Y 10 Cu 15 Al 15 .

[0059] Example 2

[0060] Based on Example 1, and different from Example 1, in step S22 of the preparation method of this embodiment, 1 wt% of the drug is added and mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder to prepare a mixed drug powder.

[0061] Recorded as experimental group 2.

[0062] Example 3

[0063] Based on Example 1, and different from Example 1, in step S22 of the preparation method of this example, 2 wt% of the drug is added and mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder to prepare a mixed drug powder.

[0064] Recorded as experimental group 3.

[0065] Example 4

[0066] Based on Example 1, and different from Example 1, in step S22 of the preparation method of this embodiment, 3 wt% of the drug is added and mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder to prepare a mixed drug powder.

[0067] Recorded as experimental group 4.

[0068] Example 5

[0069] Based on Example 1, and different from Example 1, in step S22 of the preparation method of this embodiment, 4 wt% of the drug is added and mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder to prepare a mixed drug powder.

[0070] Recorded as experimental group 5.

[0071] Corrosion resistance comparison experiment between control group 1 and experimental groups 2-5:

[0072] The preserved thermally deformed magnets prepared in each experimental group were cut into small samples by wire cutting, with the height direction being the pressing direction, i.e., the c-axis, the surface parallel to the c-axis being the side surface, and the surface perpendicular to the c-axis being the front surface.

[0073] Polish the surface to be corroded to a mirror finish and clean it. The surface to be corroded is surface A, and the side opposite to the surface to be corroded is surface B. Cover surface A with black tape, place the copper wire vertically to surface B, and secure it with conductive adhesive. Then place the sample with surface A facing down in the square groove, cover the magnet sample with a hot melt gun, and tear off the black tape after ten minutes.

[0074] A corrosion solution of 3.5wt% NaCl was prepared. A "three-electrode" CHI electrochemical analyzer was used. The open circuit potential-time curve was measured for 1800 seconds; the final open circuit potential was recorded for measuring the AC impedance spectrum, with a frequency range of 0.01Hz to 105Hz and an amplitude of 5mV; the scanning rate of the polarization curve was 1mV / s, and the scanning range was around ±0.5V of the open circuit potential. The microstructure and composition of the magnets prepared by different experimental groups were observed and analyzed using a JSM-7800F scanning electron microscope (SEM) and EDS technology.

[0075] Instructions attached Figure 2 The open circuit potential of the side of the preserved hot deformed magnet in 3.5wt% NaCl corrosion solution with different proportions of Y-Gd based alloy is shown in the attached manual. Figure 2As shown in (b), with the increase of the addition ratio of the Y-Gd-based alloy, the open circuit potential reaches a maximum value of -0.788V when the addition ratio is 2wt%. The open circuit potential gradually decreases as the addition ratio of the Y-Gd-based alloy continues to increase, but the open circuit potential of the umbilical cord mononuclear cells prepared in the experimental group is higher than that of the umbilical cord mononuclear cells in the control group. Therefore, the present invention improves the open circuit potential and the corrosion resistance of the multi-main phase magnet by adding the Y-Gd-based alloy to the mononuclear cells (generally speaking, the higher the open circuit potential, the more stable the magnet is in the corrosive medium, and the better the corrosion resistance), and the corrosion resistance of the multi-main phase magnet reaches the best when the addition ratio of the Y-Gd-based alloy is 2wt%.

[0076] From the attached drawings Figure 3 As shown in (a), the impedance spectrum size arrangement relationship is: 2wt%>3wt%>1wt%>4wt%>0wt%, that is, experimental group 3>experimental group 4>experimental group 2>experimental group 5>control group 1. The impedance spectrum radius of the experimental group magnets with added Y-Gd alloy is larger than that of the control group magnets without added Y-Gd alloy, indicating that the corrosion resistance of the magnets has been improved after adding Y-Gd alloy to mononuclear cells. When the addition amount of Y-Gd alloy is 2wt%, the impedance spectrum radius is the largest and the corrosion resistance of the magnet is the best.

[0077] Attached to the instruction manual Figure 5 As shown, the AC impedance spectrum radius of the front side of the heat-deformed multi-phase magnet prepared in the same experimental group is significantly larger than that of the side of the magnet, which indicates that the corrosion resistance of the front side of the heat-deformed umbilical cord mononuclear cell magnet prepared by the present invention is better than that of the side.

[0078] Attached to the instruction manual Figure 6 As shown in the figure, the umbilical cord mononuclear cells prepared from the control group 1 without Y-Gd alloy have the most serious corrosion. The main phase grains undergo intergranular corrosion along the mononuclear cell rare earth-rich phase, and the mononuclear cell rare earth-rich phase corrodes first. The rare earth hydrides formed by rare earth elements in the process of hydrogen absorption corrosion tend to expand in volume, while the volume of the main phase grains does not change much, causing the mononuclear cells to crack, resulting in a large number of cracks and corrosion pits, resulting in the shedding of the main phase grains. Figure 6 The magnet with 2wt% Y-Gd alloy added in (c) has the lowest corrosion degree, no obvious cracks, and the best corrosion resistance. Therefore, the present invention improves the corrosion resistance of the magnet by adding Y-Gd-based alloy at the mononuclear cells. When the addition amount of Y-Gd alloy is 2wt%, the corrosion resistance of the magnet is the best.

[0079] Comparison of side corrosion between control group 1 and experimental group 3:

[0080] Attached to the instruction manual Figure 7 As shown, Figure 7The corrosion morphology of the side of the umbilical cord mononuclear cell prepared without adding Y-Gd alloy in the control group 1 is shown. The three-point area marked by the arrows in the figure was subjected to point scanning analysis.

[0081] Attached to the instruction manual Figure 8 As shown, Figure 8 The side corrosion morphology of the umbilical cord mononuclear cells prepared by adding 2wt% Y-Gd alloy in Experimental Group 3 is shown. EDS point scanning analysis was performed on the two areas indicated by the arrows in the figure.

[0082] It can be clearly seen from the above comparison results that the magnet prepared without Y-Gd alloy is rapidly corroded in the NaCl corrosive medium, while the magnet prepared by adding Y-Gd in the present invention can effectively improve the corrosion resistance of the magnet.

[0083] In summary, by comparing the data of the test control group 1 and experimental groups 2 to 5, it is concluded that the umbilical cord mononuclear cells prepared by adding 2wt% of Y-Gd alloy drug powder in the present invention have the best therapeutic effect. The present invention uses a low-cost Y-Gd-based alloy to modify multi-main phase (Nd, Ce)-Fe-B magnet mononuclear cells to prepare umbilical cord blood-derived mesenchymal stem cells for the treatment of urinary incontinence.

[0084] Finally: The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence, characterized in that: The following steps are involved: S1: After the mother gives birth, the professional immediately cuts the umbilical cord connecting the baby and the mother, and quickly clamps the two ends of the umbilical cord with an umbilical cord clamp to prevent blood from flowing out. Then, the umbilical cord is placed in a prepared sterile bag and transferred to the laboratory as soon as possible. The same proportion of Ce-rich main phase MQP magnetic powder and Nd-rich main phase MQU-F magnetic powder are selected, as well as the drugs added to prepare mononuclear cells. The collected umbilical cord blood is anticoagulated with heparin and stored in a 4°C refrigerator to ensure that the next step of processing is carried out within 6 hours after collection; S2: The same proportion of Ce-rich main phase MQP magnetic powder and Nd-rich main phase MQU-F magnetic powder are mixed and stirred evenly with 0-4wt% of the total drug to obtain a mixed drug powder, and the umbilical cord blood and mononuclear cell separation solution are mixed in a certain proportion, put into a centrifuge tube, and subjected to density gradient centrifugation, first centrifuged at 2000×g, 4°C for 20 minutes to remove the upper plasma and platelets; S3: placing the mixed drug powder in step S2 into a centrifuge tube and storing it under vacuum to obtain a stored sample, then transferring the middle layer containing mononuclear cells into a new centrifuge tube and performing centrifugation at a higher speed, such as 10,000×g at 4° C. for 10 minutes, to further remove dead cells and cell debris; S4: placing the preserved sample into a centrifuge tube and preserving it under vacuum conditions to obtain a round cake-shaped umbilical cord mononuclear cell sample; Among them, the molecular formula of the Ce-rich main phase MQP magnetic powder in step S1 is Nd29.8Pr0.2Ga0.46Febal.Co4B0.94; the molecular formula of the Nd-rich main phase MQU-F magnetic powder is Ce23.6Nd4Pr1.9Fe69.5B; and the molecular formula of the drug added to the monocytes is Gd60Y10Cu15Al 15.

2. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 1, characterized in that: The method for preparing the drug added to the monocytes in step S1 further comprises the following steps: S11: inoculating the isolated monocyte suspension into a culture flask or culture dish pre-coated with 0.1% gelatin, and culturing in an incubator at 37° C. and 5% CO 2 ; S12: Umbilical cord mesenchymal stem cells (MSCs) will gradually adhere to the bottom of the culture bottle during the culture process due to their characteristic of adherent growth, while other non-adherent cells will be gradually removed by changing the medium; S13: Usually, the medium needs to be changed three times after the first adhesion, with an interval of 3-4 days between each change, to remove non-adherent cells and impurities.

3. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 2, characterized in that: The method for preparing the mixed drug powder in step S2 further comprises the following steps: S21: first weigh 50 parts by weight of Ce-rich main phase MQP magnetic powder and 50 parts by weight of Nd-rich main phase MQU-F magnetic powder; S22: Then, the drug with an addition amount of 1wt%, 2wt%, 3wt% or 4wt% is uniformly mixed with the Ce-rich main phase MQP magnetic powder and the Nd-rich main phase MQU-F magnetic powder to prepare mixed drug powders containing different contents of alloy substances.

4. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 3, characterized in that: In step S3, the method for preserving the sample further comprises the following steps: S31: first pour the mixed drug powder into a centrifuge tube with a diameter of φ20 for storage; S32: The mixed drug powder is then placed in the center of the furnace of the centrifuge tube, and after vacuuming, the heating and pressurization procedures are started simultaneously.

5. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 4, characterized in that: The method for preserving and molding the umbilical cord mononuclear cell sample in step S4 further comprises the following steps: S41: first take out the dense precursor sample in step S3 and put it into a φ36 centrifuge tube; S42: Then, the mixed drug powder is placed in the middle of the furnace of the centrifuge tube, and after vacuuming, it is heated and started. α-MEM culture medium containing 10% fetal bovine serum (FBS) is used, and antibiotics such as penicillin and streptomycin are added to maintain the cells to proliferate under the best growth conditions. The cell growth is observed every day, and subculture is performed in a timely manner to maintain cell viability. MSCs are further purified by flow cytometry or immunomagnetic beads to remove residual impurity cells and improve cell purity. When the number of cells reaches a certain scale, some cells can be frozen for subsequent use. The use of amorphous physical vitrification method for cell freezing can ensure that the cells maintain complete morphology and function during the storage process.

6. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 5, characterized in that: In step S4, the deformation amount of the Cerium Iron Boron magnet sample is 70%, and the outer circle size is φ36 and the height is 8mm in a round cake-shaped structure.

7. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 6, characterized in that: The temperature parameters of the saving process in step S32 are set as follows: heating to 300°C at an output power of 30W, then heating to 550°C at an output power of 50W, keeping warm for 10 minutes, then continuing to heat to 680°C at this power, and finally keeping warm for 30 minutes.

8. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 7, characterized in that: In step S32, the process pressure parameter settings are saved: pressurizing to 60 MPa, and then maintaining the pressure for 30 minutes.

9. The preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to claim 8, characterized in that: The saved process parameters in step S42 are set as follows: heating to 300°C at an output power of 30W, then heating to 650°C at an output power of 50W, keeping warm for 10 minutes, then continuing to heat to 750°C at this power, and finally keeping warm for 20 minutes. When the temperature reaches 750°C, the pressure is started, and it takes 20 minutes (deformation rate is 0.001s-1) to increase the pressure to 90MPa.

10. Umbilical cord mononuclear cells prepared according to the preparation method and application method of mesenchymal stem cells derived from umbilical cord blood for treating urinary incontinence according to any one of claims 1 to 9, characterized in that: The atomic percentage of the NdxCeyFe100-xyz-nBzMn magnetic powder component is NdxCeyFe100-xyz-nBzMn, wherein 12≤x+y≤16, 5.5≤z≤7, 0.1≤n≤5, and M is one or more of the elements Co, Ni, Al, Cu, Ga, Mg, Zn, Nb, Zr, Ti, Mo, W, and V.

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