Application of CD146 + umbilical cord mesenchymal stem cell subpopulation in preparation of medicine for preventing and treating immune thrombocytopenia and product

Abstract

The invention discloses application of CD146 + umbilical cord mesenchymal stem cell subpopulation in preparation of drugs for preventing and treating immune thrombocytopenia and a product, and relates to the technical field of biology. The invention provides an application of a CD146 + mesenchymal stem cell subset in preparation of a medicine for treating or preventing immune thrombocytopenia. The CD146 < + > mesenchymal stem cell subpopulation can increase the number of platelets of ITP patients, regulate the CD4 < + > / CD8 < + > ratio of the ITP patients and remarkably reduce the MDSC proportion, so that the in-vivo immune state is changed.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology, in particular to the application of CD146+umbilical cord mesenchymal stem cell subpopulation in the preparation of drugs for preventing and treating immune thrombocytopenia. BACKGROUND

[0002] Primary immune thrombocytopenia (ITP), also known as idiopathic thrombocytopenic purpura, is an acquired autoimmune disease. It is the most common bleeding disease caused by a decrease in platelet count. It is generally recognized that most ITP is caused by immune-mediated autoantibody sensitized platelets being excessively destroyed by the mononuclear macrophage system. There is also a new view that it is immune-mediated damage to megakaryocytes or inhibition of megakaryocyte release of platelets, resulting in insufficient platelet production in patients. ITP is characterized by extensive skin and mucosal and internal organ bleeding, thrombocytopenia, and other clinical features.

[0003] Mesenchymal stem cell (MSC) therapy has made significant progress in the study of its role in multiple autoimmune diseases. It has low immunogenicity between biological individuals, can induce immune tolerance and reduce inflammatory damage, but mesenchymal stem cells have heterogeneity at multiple levels, including differences between donors, tissues, subpopulations, and different batches. In addition, the proportion of subpopulation components in mesenchymal stem cell products developed by different processes varies greatly. CD146 is a membrane protein and one of the markers of mesenchymal stem cell subtypes. Previous studies on the therapeutic effects of human umbilical cord CD146+MSCs found that they can have therapeutic effects on ischemic stroke, premature ovarian failure, and acute-on-chronic liver failure models through immune regulation and angiogenesis.

[0004] A preparation method of genetically modified mesenchymal stem cell-derived exovesicles and application thereof are disclosed in Chinese patent CN119220498A. The preparation method comprises the following steps: S1, taking microcarriers and culture medium in a mass ratio of 1:100, and placing the microcarriers and culture medium into a glass flask to prepare a culture bottle; S2, placing mesenchymal stem cells stably overexpressing PD-L1 and ICAM-1 molecules into the culture bottle to obtain a three-dimensional suspension culture system of mesenchymal stem cells; S3, placing the three-dimensional suspension culture system of mesenchymal stem cells on a stirrer with stirring parameters of 40-60 rpm, stirring for 15-20 min, and intermittent duration of 2-3 h; S4, collecting the culture solution every 48-72 h during the culture process, and continuously culturing for 28-30 days to obtain total culture solution; and S5, performing ultrafiltration concentration and high-speed centrifugation on the obtained total culture solution to obtain mesenchymal stem cell-derived exovesicles expressing PD-L1 and ICAM-1. After intravenous injection of the mesenchymal stem cell-derived exovesicles overexpressing PD-L1 and ICAM-1 genes into a mouse model of immune thrombocytopenia, the number of platelets can be significantly increased, and subcutaneous purpura disappears. However, the genetically modified mesenchymal stem cell-derived exovesicles increase the safety risk of treatment.

[0005] The current clinical treatment bottleneck is that the long-term efficacy of drugs is poor, and the adverse reactions are many, and there is still no ideal prevention and treatment method for refractory ITP patients who are ineffective for hormone and splenectomy treatment. Therefore, there is a lack of more effective drug development strategies with safety and avoiding side effects. SUMMARY

[0006] The purpose of the present application is to provide the application of CD146+umbilical cord mesenchymal stem cell subpopulation in the preparation of drugs for preventing and treating immune thrombocytopenia and products.

[0007] To achieve the above-mentioned purpose of the application, the technical solutions of the present application are as follows: On the one hand, the present application provides the application of CD146+mesenchymal stem cell subpopulation in the preparation of drugs for treating or preventing immune thrombocytopenia.

[0008] Specifically, the drug has at least one of the following effects: (1) increasing the number of platelets in patients with immune thrombocytopenia; (2) regulating the ratio of CD4+ / CD8+; (3) regulating the ratio of CD11b / Gr-1.

[0009] Further, the drug exerts the effect of treating immune thrombocytopenia through immune regulation.

[0010] Still further, the immune regulation effect comprises: (1) reducing the proportion of GR-1; (2) increasing the proportion of CD11b / Gr-1; (3) inhibiting the reaction of T cells, NK cells and B cells.

[0011] Specifically, the CD146+mesenchymal stem cell subpopulation comprises a CD146+mesenchymal stem cell subpopulation separated from Wharton's jelly of umbilical cord.

[0012] Further, the mesenchymal stem cells comprise, but are not limited to, umbilical cord-derived mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, placenta-derived mesenchymal stem cells, fat-derived mesenchymal stem cells, dental pulp-derived mesenchymal stem cells, and uterine blood-derived mesenchymal stem cells.

[0013] Still further, the mesenchymal stem cells are umbilical cord-derived mesenchymal stem cells.

[0014] Further, the preparation method of the CD146+umbilical cord mesenchymal stem cell subpopulation is Wharton's jelly enzyme digestion.

[0015] Specifically, the CD146+mesenchymal stem cell subpopulation highly expresses CD73, CD90 and CD105.

[0016] Specifically, the CD146+mesenchymal stem cell subpopulation does not express CD34 and CD45.

[0017] Specifically, the expression rate of CD146 in the CD146+mesenchymal stem cell subpopulation is not less than 80%.

[0018] According to some embodiments of the present application, the expression rate of CD146 in the CD146+mesenchymal stem cell subpopulation can be 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, and any intermediate point value.

[0019] Further, the expression rate of CD146 in the CD146+mesenchymal stem cell subpopulation is not less than 90%.

[0020] Further, the expression rate of CD146 in the CD146+mesenchymal stem cell subpopulation is not less than 95%.

[0021] Further, the expression rate of CD146 in the CD146+mesenchymal stem cell subpopulation is not less than 97%.

[0022] Specifically, the content of the CD146+ mesenchymal stem cell subpopulation in the medicine is not less than 2×10 5 / 100 μL.

[0023] Further, the content of the CD146+ mesenchymal stem cell subpopulation in the medicine is not less than 2.5×10 5 / 100 μL.

[0024] Still further, the content of the CD146+ mesenchymal stem cell subpopulation in the medicine is 2.5×10 5 / 100 μL or 5×10 5 / 100 μL.

[0025] Specifically, the medicine further comprises a pharmaceutically acceptable carrier.

[0026] Further, the pharmaceutically acceptable carrier is selected from one or more of excipients, buffers, emulsifiers, coloring agents, anti-adhesion agents, penetration enhancers, plasticizers, surfactants, antifoaming agents, thickening agents, inclusion agents, absorbents, humectants, propellants, solubilizers, cosolvents, pH adjustors, binders, disintegrants, fillers, lubricants, wetting agents, integrating agents, osmotic pressure adjustors, stabilizers, glidants, flavoring agents, preservatives, foaming agents, suspending agents, coating materials, fragrances, diluents, flocculants, deflocculants, filter aids.

[0027] Specifically, the dosage form of the medicine is a tablet, a powder, a granule, a capsule, a suspension, an oil, a spray, an aerosol or an injection.

[0028] Further, the dosage form of the medicine is an injection.

[0029] Specifically, the injection includes but is not limited to an intravenous injection, an intramuscular injection, a subcutaneous injection, an intradermal injection, a cavity injection.

[0030] In another aspect, the present application provides a product for treating or preventing immune thrombocytopenia, wherein the product comprises a CD146+ mesenchymal stem cell subpopulation.

[0031] Specifically, the product is a medicine.

[0032] Specifically, the medicine has at least one of the following effects: (1) increasing the number of platelets of a patient with immune thrombocytopenia; (2) regulating the ratio of CD4+ / CD8+; (3) regulating the ratio of CD11b / Gr-1.

[0033] Specifically, the CD146+ mesenchymal stem cell subpopulation comprises CD146+ mesenchymal stem cells separated from umbilical cord Wharton's jelly.

[0034] Further, the expression rate of CD146 in the CD146+ mesenchymal stem cell subpopulation is not less than 80%.

[0035] Further, the expression rate of CD146 in the CD146+ mesenchymal stem cell subpopulation is not less than 90%.

[0036] Further, the expression rate of CD146 in the CD146+ mesenchymal stem cell subpopulation is not less than 95%.

[0037] Further, the expression rate of CD146 in the CD146+ mesenchymal stem cell subpopulation is not less than 97%.

[0038] Specifically, the content of the CD146+ mesenchymal stem cell subpopulation in the drug is not less than 2×10 5 / 100 μL.

[0039] Further, the content of the CD146+ mesenchymal stem cell subpopulation in the drug is not less than 2.5×10 5 / 100 μL.

[0040] Still further, the content of the CD146+ mesenchymal stem cell subpopulation in the drug is 2.5×10 5 / 100 μL or 5×10 5 / 100 μL.

[0041] Specifically, the drug further comprises a pharmaceutically acceptable carrier.

[0042] Specifically, the dosage form of the drug comprises tablets, powders, granules, capsules, suspensions, oils, sprays, aerosols, injections.

[0043] Further, the dosage form of the drug is injection.

[0044] The beneficial effects of the present application are: The application discloses a treatment of immune thrombocytopenia using a non-genetically modified CD146+mesenchymal stem cell subpopulation. The application provides a treatment of immune thrombocytopenia using a CD146+mesenchymal stem cell subpopulation, and finds that the CD146+mesenchymal stem cell subpopulation can increase the platelet count of ITP mice, increase the CD4+ / CD8+ratio of ITP mice (CD3+CD4+ / CD3+CD8+represents the ratio of T helper cells / T suppressor cells), and significantly reduce the proportion of MDSC, thereby changing the immune state in vivo. It is indicated that the CD146+mesenchymal stem cell subpopulation mainly plays a therapeutic role in ITP mice through immune regulation. BRIEF DESCRIPTION OF DRAWINGS

[0045] Figures 1-2 The cell phenotype of the CD146+MSCs is that the expression rates of CD34 and CD45 (marked as Neg) are less than 2%, the expression rates of CD73, CD90 and CD105 are greater than 95%, and the expression rate of CD146% is 97.3%.

[0046] Figure 3 The platelet count of the CD146+MSCs after treatment of immune thrombocytopenia mice is shown in the figure (n=5 / 6, *P<0.05, ***P<0.001).

[0047] Figure 4 The effect of the CD146+MSCs on CD4+ / CD8+T cells after treatment of immune thrombocytopenia mice is shown in the figure (n=5 / 6, *P<0.05).

[0048] Figure 5 The effect of the CD146+MSCs on CD4+ / CD8+T cells after treatment of immune thrombocytopenia mice in the control group.

[0049] Figure 6 The effect of the CD146+MSCs on CD4+ / CD8+T cells after treatment of immune thrombocytopenia mice in the model group.

[0050] Figure 7 The effect of the CD146+MSCs on CD4+ / CD8+T cells after treatment of immune thrombocytopenia mice in the low-dose group.

[0051] Figure 8 The effect of the CD146+MSCs on CD4+ / CD8+T cells after treatment of immune thrombocytopenia mice in the medium-dose group.

[0052] Figure 9 The effect of the CD146+MSCs on CD4+ / CD8+T cells after treatment of immune thrombocytopenia mice in the high-dose group.

[0053] Figures 10-13 For the influence of CD146+MSCs on MDSC cells after treating immune thrombocytopenia mice, (n=5 / 6, *P<0.05).

[0054] Figures 14-18 For the flow chart and statistical chart of CD146 expression level detection of different umbilical cord-derived mesenchymal stem cells, (n=3 / 5, **P<0.01).

[0055] Figure 19 For the T cell inhibition situation of CD146 low expression and high expression MSCs (**P<0.01).

[0056] Figure 20 For the comparison of iNOS, TNF-α and IL-6 content detection of 6h LPS pretreatment group (**P<0.01). DETAILED DESCRIPTION

[0057] In order to make the technical means, creative characteristics, purposes and effects realized by the present application easy to understand, the following specific embodiments are further illustrated. The following embodiments are preferred embodiments of the present application, but not all. Based on the embodiments in the embodiments, other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application. In the following embodiments, if not otherwise specified, the operation method used is a conventional operation method, the equipment used is a conventional equipment, and the equipment materials used in each embodiment are the same.

[0058] Example 1 1. Source and subculture method of CD146+mesenchymal stem cell subpopulation 1) The first generation of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) were prepared and provided by Beijing Sanhuailikang Cell Technology Co., Ltd. The Wharton's jelly part of the umbilical cord was mechanically separated, cut into 1-3 mm small pieces, and then added with 15 mL of collagenase I (Sigma, USA) with a final concentration of 0.5% for digestion to obtain P0 generation cells, which were further subcultured (P) at 37°C, 5% CO2 to obtain P1 generation cells. The cells were collected with 0.5% trypsin-EDTA (Gibco, Australia), counted, and then reseeded into a 100 mm cell culture dish at a density of 20,000 cells / cm2. 3 1) The first generation of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) were prepared and provided by Beijing Sanhuailikang Cell Technology Co., Ltd. The Wharton's jelly part of the umbilical cord was mechanically separated, cut into 1-3 mm small pieces, and then added with 15 mL of collagenase I (Sigma, USA) with a final concentration of 0.5% for digestion to obtain P0 generation cells, which were further subcultured (P) at 37°C, 5% CO2 to obtain P1 generation cells. The cells were collected with 0.5% trypsin-EDTA (Gibco, Australia), counted, and then reseeded into a 100 mm cell culture dish at a density of 20,000 cells / cm2.

[0059] 2) Phenotypic characterization Flow cytometry (Beckman, cytoflexs) was used to detect the percentage of CD146 positive cells. Cells were divided into two tubes, 4 x 10 5 cells / tube, and anti-bodies: CD45-FITC, CD34-FITC, CD90-PerCP, CD73-PE, CD105-APC (Sino Biological) and CD146-FITC (BioLegend) were added respectively at 4°C for 30 min. After washing twice with PBS, the percentage of cells was detected by flow cytometry.

[0060] 3) Preparation of the preparation After digestion, hUC-MSCs cells were resuspended in a solution of normal saline (Shijiazhuang Four Pharmaceutical Group) containing 5% human blood albumin (Jet Bern).

[0061] 2 Preparation of guinea pig anti-mouse platelet serum (GP-APS) 2.1 Preparation of mouse platelets (APS) 1) BALB / C mice were hearted to collect whole blood (ethylenediaminetetraacetic acid, i.e. EDTA anticoagulation), and centrifuged at 1200 rpm for 10 minutes; 2) The upper platelet-rich plasma was centrifuged at 14000 rpm for 10 minutes, and the supernatant was discarded; 3) The precipitate was washed with platelet washing solution for 3 times, and the precipitate was resuspended with platelet suspension, and the platelet concentration was adjusted to 1 x 10 10 / L.

[0062] 2.2 Preparation of guinea pig anti-mouse platelet serum (GP-APS) 1) The prepared platelets were mixed with an equal volume of complete Freund's adjuvant to prepare a suspension, and were injected subcutaneously on the footpads, back and subcutaneously of guinea pigs once (4 points per, 100 μL per point); 2) On the 7th, 14th and 28th day after the first injection, the platelet suspension mixed with an equal volume of incomplete Freund's adjuvant was injected subcutaneously on the footpads, back and subcutaneously of guinea pigs (4 points per, 100 μL per point); 3) On the 35th day, the whole blood without anticoagulation was centrifuged at 560g for 10 minutes, and the supernatant (GP-APS) was aliquoted and stored at -20°C.

[0063] 3 Construction of mouse immune thrombocytopenia model (1) The APS was taken out from -20°C and placed in a 56°C water bath for 30 min, and was adsorbed with an equal amount of BALB / C mouse red blood cells for at least twice, and was diluted with normal saline to 1:4 GP-APS.

[0064] (2) After adaptive feeding of mice, GP-APS (200 μL) was injected intraperitoneally, and drug injection intervention was performed on the 2nd, 4th, and 6th days after injection.

[0065] 4 Mesenchymal stem cell treatment of mouse immune thrombocytopenia 4.1 Grouping The solvent was a physiological saline solution containing 5% human blood albumin.

[0066] 1) Control group (n=10): normal feeding, tail vein injection of solvent (200 μL); 2) Model group (n=10): intraperitoneal injection of GP-APS + tail vein injection of solvent (200 μL); 3) Low-dose group (n=10): intraperitoneal injection of GP-APS + tail vein injection of stem cells (2×10 5 / each time, 200 μL); 4) Medium-dose group (n=10): intraperitoneal injection of GP-APS + tail vein injection of stem cells (5×10 5 / each time, 200 μL); 5) High-dose group (n=10): intraperitoneal injection of GP-APS + tail vein injection of stem cells (1×10 6 / each time, 200 μL).

[0067] 4.2 Detection items 1) Before GP-APS injection, once every 2 days after GP-APS injection, whole blood samples were taken for blood routine test (6 per group per time); 2) On the 5th day after GP-APS injection (after the 2nd intervention), for flow detection (5 per group per time) Flow cytometry was used to detect lymphocytes (CD3 / CD4 / CD8), myeloid-derived suppressor cells (CD11b / Gr-1) in peripheral blood on the 5th day after GP-APS injection (after the 2nd intervention).

[0068] 4.3 Experimental results 4.3.1 Characteristics of umbilical cord CD146+mesenchymal stem cells The CD146+subgroup highly expressed the human umbilical cord mesenchymal stem cell-related markers CD73, CD90, and CD105; did not express the hematopoietic stem cell-related markers CD34 and CD45, and the expression rate of CD146+was 97.4%. See Figures 1-2 .

[0069] 4.3.2 Platelet count The platelet count of the model group and the low, medium and high dose groups of the immune thrombocytopenia mice was significantly reduced on the second day after the model was established, indicating that the model was successfully established; on the 16th and 18th days after drug treatment, the platelet count of the high dose group was significantly increased, and there was a significant difference with the model group (n=5 / 6, *P<0.05, ***P<0.01), as shown in Figure 3 .

[0070] 4.3.3 Changes in peripheral blood immune cells 1) CD3+CD4+ / CD3+CD8+ represents the ratio of T helper cells / T suppressor cells, and the detection results of lymphocytes (CD3 / CD4 / CD8) in the peripheral blood of ITP mice on the 5th day after GP-APS injection (after the second intervention) showed that the CD8+ cells of the high dose group were significantly decreased, and the CD4+ / CD8+ ratio was increased; (n=5 / 6, *P<0.05), as shown in Figures 4-9 .

[0071] 2) GR-1 is a marker of myeloid-derived suppressor cells (MDSC), and after treatment of ITP mice, the proportion of GR-1 in the peripheral blood of the high dose group was significantly reduced, and the proportion of myeloid-derived suppressor cells (CD11b / Gr-1) was increased. MDSC is a heterogeneous cell population derived from bone marrow cells, which is characterized by the ability to suppress the response of T cells, NK cells and B cells, thereby changing the immune status in the body. It is indicated that the CD146+ mesenchymal stem cell subpopulation mainly plays a therapeutic role in ITP mice through immune regulation Figures 10-13 .

[0072] Comparative Example In vitro immune regulation of MSCs with different CD146 expression levels 1. Study on the inhibition of T cells by MSCs with different CD146 expression levels (1) CD146 expression detection of two groups (8 strains) of MSC cells All cells were recovered in turn, and 1×10 6 Cells were added with 2 μL / tube of APC-CD146 dye for CD146 expression detection.

[0073] Results: The recovered cells were labeled with CD146 antibody, and the flow cytometry detection results are shown in Figures 14-18 Table 1 summarizes the MSCs of the CD146 low expression group (n=3, MSC-1, 2, 3) and the high expression group (n=5, MSC-4, 5, 6, 7, 8), and the results show that the two selected groups can significantly distinguish the expression level of CD146 (P<0.01).

[0074] Table 1 CD146 expression level of different umbilical cord-derived mesenchymal stem cells

[0075] (2) Two groups (8 strains) of MSC cells on T cell inhibition Take the number of cells about 2 x 10 5 MSC, add complete medium containing mitomycin C (10 μg / ml), 37°C, 5% CO2 incubator for 30 min; sodium chloride injection wash once, using immune cell culture medium to adjust the cell concentration to 3 x 10 5 / ml, 50 μl / well to 96-well plate; Luci-T cell (Beijing San Yilikon Cell Technology Co., Ltd.) was recovered, and the Luci-T cell concentration was adjusted to 1 x 10 6 / ml, 50 μl / well to 96-well plate, and placed in a 37°C, 5% CO2 incubator; continuous culture for 4d, 50 μl of fluorescent substrate was added to each well, and the fluorescence intensity was detected.

[0076] Table 2 CD146 low expression and high expression group on T cell inhibition rate

[0077] The results are shown in Figure 19 Due to the individual differences of umbilical cord, different umbilical cord-derived mesenchymal stem cells have different effects on T cell inhibition. MSC (MSC-4, 5, 6, 7, 8) with high CD146 expression have significantly higher immune inhibition effect on T cells than MSC with low CD146 expression (P<0.01).

[0078] 2 Comparison of CD146+, -MSC exosomes on the regulation of inflammatory factors (1) Experimental grouping N9 cells are mouse microglial cell lines, which are cultured and expanded according to the culture conditions of IMDM + 5% FBS + 1% PS + 50 uM β-mercaptoethanol. The cells are resuspended with N9 complete culture medium, and the density is adjusted to 1 x 10 5 / mL, and the cells are inoculated in 24-well plates with a culture volume of 1 mL. The culture groups are as follows: Table 3 Experimental grouping

[0079] (2) Preparation method of exosomes ① Collect CD146+MSC, CD146-MSC cells and culture supernatant, centrifuge at 300 g for 10 minutes, remove cells and large particles; ② Keep the supernatant in ①, centrifuge at 2000 g for 10 minutes at 4°C, remove cell debris; ③ Keep the supernatant of ②, centrifuge at 2000 g for 30 minutes at 4℃, remove the large membrane bubbles; ④ Keep the supernatant of ③, centrifuge at 100000 g for 75 minutes at 4℃, further purification; ⑤ Remove most of the supernatant of ④, keep 5 mL, reduce the loss of exosome precipitation; ⑥ Centrifuge at 100000 g for 75 minutes at 4℃, remove all supernatant, and resuspend the precipitate with 1 mL of PBS.

[0080] (3) Cell pretreatment and LPS stimulation According to the grouping in Table 3, the cells were inoculated, pretreated with LPS for 6 hours according to the grouping, and then treated with exosomes for 24 hours.

[0081] (4) Factor detection The cell supernatant was taken into a centrifuge tube, centrifuged according to the sample processing requirements of different factor detection kits, aliquoted, and stored at -20℃. Mouse IL-6 factor (Link Biotech, item number EK206), TNF-a (Biolegend, item number 430904), and iNOS factor (Jianglai Biotech, item number JL20675) detection were set up with 1 replicate well, and each sample had a total of 2 detection wells, which were performed according to the relevant factor detection instructions.

[0082] Results: The LPS group successfully stimulated the increased expression of IL-6, iNOS, and TNF-a factors. After adding high and low concentrations of CD146+ / - exosomes, the contents of iNOS, TNF-a, and IL-6 decreased, among which the CD146+MSC exosomes had a significant effect, and had a concentration effect, that is, the effect was better at high concentration and decreased at low concentration (see Figure 20 ).

[0083] This experiment can prove that the anti-inflammatory effect of CD146+MSC exosomes is more significant than that of CD146-MSC exosomes.

[0084] (1) iNOS content Table 4 iNOS content (IU / mL)

[0085] (2) TNF-a content Table 5 TNF-a content (pg / mL)

[0086] (3) IL-6 content Table 6 IL-6 content (pg / mL)

[0087] In summary, MSCs with high CD146 expression levels have superior immune regulation and anti-inflammatory effects, and CD146+MSCs can exert therapeutic effects on ITP disease through immune regulation.

[0088] The above merely describes preferred embodiments of the present application and is not intended to limit the present application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Application of CD146+ mesenchymal stem cell subsets in the preparation of drugs for the treatment or prevention of immune thrombocytopenic purpura.

2. The application according to claim 1, characterized in that, The drug has at least one of the following effects: (1) Increase platelet count in patients with immune thrombocytopenic purpura; (2) Adjusting the CD4+ / CD8+ ratio; (3) Adjust the CD11b / Gr-1 ratio.

3. The application according to claim 1, characterized in that, The CD146+ mesenchymal stem cell subpopulation includes CD146+ mesenchymal stem cells isolated from Wharton's jelly via the umbilical cord.

4. The application according to claim 1, characterized in that, The mesenchymal stem cells mentioned include umbilical cord-derived mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, placental-derived mesenchymal stem cells, adipose-derived mesenchymal stem cells, dental pulp-derived mesenchymal stem cells, and uterine blood-derived mesenchymal stem cells.

5. The application according to claim 4, characterized in that, The mesenchymal stem cells mentioned are umbilical cord-derived mesenchymal stem cells.

6. The application according to claim 1, characterized in that, The expression rate of CD146 in the CD146+ mesenchymal stem cell subset is not less than 80%.

7. The application according to claim 1, characterized in that, The drug contains at least 2 × 10⁻⁶ CD146+ mesenchymal stem cell subsets. 5 / 100μL.

8. The application according to any one of claims 1-7, characterized in that, The drug also includes a pharmaceutically acceptable carrier.

9. The application according to any one of claims 1-7, characterized in that, The dosage form of the drug is tablet, powder, granule, capsule, suspension, oil, spray, aerosol or injection.

10. A product for treating or preventing immune thrombocytopenic purpura, characterized in that, The product includes a CD146+ mesenchymal stem cell subpopulation.

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