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CD24 positive expression cell in urine as well as preparation method and application of CD24 positive expression cell

A cell and urine technology, which is applied in the field of CD24 positive expression cells and its preparation, can solve the problems of restricting the application of muscle tissue regeneration, achieve the effect of muscle regeneration and repair, avoid immune rejection, and reduce the symptoms of inflammatory response

Active Publication Date: 2022-05-17
广东明珠生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, with the development of stem cells and tissue engineering methods, new methods have been provided for the treatment of muscle injuries. The implantation of tissue engineering materials can assist in the treatment of large-volume muscle defects. Due to the technical barriers of activity and biocompatibility, implantable materials will cause a large rejection reaction in the body, which greatly limits the application of muscle tissue regeneration; and stem cell therapy is due to cell source / subgroup classification and growth The difference in muscle differentiation function, the therapeutic effect on muscle injury repair is still in the stage of further research

Method used

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  • CD24 positive expression cell in urine as well as preparation method and application of CD24 positive expression cell
  • CD24 positive expression cell in urine as well as preparation method and application of CD24 positive expression cell
  • CD24 positive expression cell in urine as well as preparation method and application of CD24 positive expression cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This embodiment is the separation and expansion culture method of CD24 positive expressing cells in urine:

[0033] (1) Source of urine: from the patient who is taking the drug, collect 100-300ml of the patient’s own urine into a sterile container, divide the urine into 50ml centrifuge tubes, centrifuge at 400g for 5 minutes, and collect the cell sediment;

[0034] (2) Use 5ml of KSFM culture medium to resuspend the cell pellet obtained in step (1), transfer to a T25 culture flask for primary culture at 37°C;

[0035] (3) For the urine cells that appeared after 3-4 days of primary culture in step (2), the medium was changed once every 2 days, and when the degree of cell fusion reached 90%, the cells were subcultured and expanded using KSFM culture medium. After the cells are expanded to the required number, they are frozen or used to obtain subcultured cells;

[0036] (4) Collect the subcultured cells in step (3), and use the flow sorting magnetic bead kit of Meitiani ...

Embodiment 2

[0044] This example is for the expression identification of the CD24 positive expression cells isolated and obtained in Example 1, and the specific identification method is as follows:

[0045] (1) Collect the cells of each group respectively, extract the RNA samples of the cells, and detect the expression of the CD24 gene by the cell population of each group by PCR reaction;

[0046] (2) Cells in each group were collected, cell protein samples were extracted, and the expression of CD24 protein in each group of cell populations was detected by western blot reaction and CD24 antibody.

[0047] For the final results see Figure 4 , Figure 4 A is the expression level of CD24 gene for CD24 negative cell population, ungrouped cells and CD24 positive cell population; Figure 4 B is the western blot reaction result graph of CD24 negative cell population, ungrouped cells and CD24 positive cell population for the expression level of CD24 protein, as can be seen from the figure, in the...

Embodiment 3

[0049] This example is the application of the CD24 positive expression cells obtained in Example 1, which is mainly used in muscle regeneration and repair. The specific experimental method includes the following steps:

[0050] (1) Establishment of rat muscle injury model:

[0051] Rat muscle injury model was induced by injecting 0.2 mg / ml CTX (cardiotoxin) at the middle part of rat tibialis anterior muscle.

[0052] (2) The rats of the rat muscle injury model are equally divided into two groups, wherein one group is the experimental group, and the other group is the control group:

[0053] The experimental group was treated with CD24 positive expressing cell injection. The specific method was as follows: the CD24 positive expressing cells obtained in Example 1 were resuspended in normal saline for injection, and prepared into CD24 positive expressing cell injection. 5 -1×10 8 Cells / ml was injected into the muscle injury site of rats for in situ repair treatment.

[0054] T...

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Abstract

The invention relates to the field of biological medicine, in particular to a CD24 positive expression cell in urine and a preparation method and application thereof. According to the application, cells with CD24 positive expression in urine-derived cells are found and screened from urine of a muscle injury patient, and the cells with CD24 positive expression in the urine cells can be subjected to myogenic differentiation under an in-vitro induction condition and are fused with muscle cells to form fused muscular tubules; it is verified that after CD24 positive expression cells in urine cells are transplanted into animal bodies, inflammatory reaction symptoms can be relieved at muscle injury parts, muscle regeneration and repair can be achieved, the CD24 positive expression cells can be used for preparing related medicine for repairing muscle injury treatment, the cells for treatment are derived from patient bodies and have the characteristic of individualized precise treatment, and the CD24 positive expression cells can be used for preparing the related medicine for repairing muscle injury treatment. The autologous cell treatment of the patient can be quickly realized, and the immunological rejection of transplantation is avoided.

Description

【Technical field】 [0001] The invention relates to the field of biomedicine, in particular to CD24-positive expression cells in urine and a preparation method and application thereof. 【Background technique】 [0002] Muscle damage and muscle loss caused by external trauma, aging and other factors can cause symptoms such as stress urinary incontinence and skeletal muscle defects, which will eventually lead to difficulty in movement and mobility of patients, which will cause great trouble to patients' daily life and social activities . At present, the clinical treatment for this kind of muscle injury is mainly limited to the patient's own rehabilitation physical therapy and some chemical drug therapy. In addition to the widely used physical rehabilitation and chemical therapy, engineering material transplantation therapy and cell therapy are also applied in a small amount to large-volume muscle defects. Specific drugs. [0003] Stress urinary incontinence, large-volume skelet...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/22A61P21/00C12N5/071
CPCA61K35/22A61P21/00C12N5/0684C12N2509/00
Inventor 易桦林谢水林邹芬
Owner 广东明珠生物技术有限公司
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