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Obtaining method and application of exosomes derived from human urine cells

A technology of human urine and exosomes, applied in the field of biomedicine

Active Publication Date: 2019-07-12
上海艾棵颂生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the method of obtaining exosomes from urine-derived cells and using them to repair tissue damage and treat diseases has not been reported

Method used

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  • Obtaining method and application of exosomes derived from human urine cells
  • Obtaining method and application of exosomes derived from human urine cells
  • Obtaining method and application of exosomes derived from human urine cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Repair of skin defects with exosomes compounded with human urine-derived cells

[0036] The skin defect model of New Zealand white rabbits was established, and they were randomly divided into control group and treatment group. In the treatment group, exosomes from human urine-derived cells were locally injected into the skin defect, and the exosomes of human urine-derived cells were analyzed from gross observation, tissue section, and immunohistochemistry at 4, 8, 12, and 14 days respectively. Effects of exudates on wound healing from skin defects in New Zealand white rabbits. It was found that the wound healing time of the treatment group was shorter than that of the control group. The wound healing time of the New Zealand white rabbits in the two groups was 16 days and 20 days respectively. Days 8, 12, and 14 were significantly smaller than those of the control group (p Figure 4 , Figure 5 , Image 6 ). The results confirmed that exosomes from human urine-derived...

Embodiment 2

[0038] Tissue-engineered bone repairing large bone defects with exosomes compounded with human urine-derived cells

[0039] A 5mm femoral defect model of 30 SD rats was made and randomly divided into groups A and B: group A, implanted with β-TCP alone; group B, implanted with β-TCP compounded with exosomes from human urine-derived cells. The animals were sacrificed in batches at 4, 8, and 12 weeks after operation, and the general condition, general observation of the defect area, X-ray, micro-CT, histological staining analysis, and immunohistochemical staining were performed to detect the fracture healing, and the bone defect repair rate of each group was compared. Effect. The results showed that the bone defect in the exosome material group compounded with human urine-derived cells could heal completely, and it could significantly promote the repair of large bone defects.

Embodiment 3

[0041] Hydrogels Composite with Exosomes from Human Urine-Derived Cells Repair Cartilage Defects

[0042] 30 New Zealand white rabbits were made with hind leg femoral condyle articular defect models, the defect diameter was 4.5mm, and the depth was 3mm, and they were randomly divided into groups A and B: group A, implanted with hydrogel alone; group B, implanted with composite Hydrogel of exosomes from human urine-derived cells. Animals were sacrificed in batches at 4, 12, and 18 weeks after operation, and the general condition, general observation of the defect area, X-ray, micro-CT, histological staining analysis, and immunohistochemical staining were performed to detect fracture healing, and the bone defect repair rate of each group was compared. Effect. The results showed that the cartilage defect in the exosome material group compounded with human urine-derived cells could heal completely and significantly promote the repair of cartilage defect.

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Abstract

The invention provides an acquisition method for exosomes derived from human urinary cells. The acquisition method includes the steps that firstly, human urine source cells are separately cultured, a culture medium is collected, the culture medium of the human urine source cells is filtered through a 0.22-micrometer filter membrane, and then large cell fragments and other impurities are removed; then an organelle is centrifugally removed, and supernatant is reserved; a membrane capable of intercepting 100KD molecular weight is used, the exosomes in the supernatant are centrifugally intercepted, after interception, PBS is used for eluting the membrane, and an exosome concentrated solution is obtained. The exosomes can be used for preparing medicine with the effects of resisting apoptosis, promoting angiogenesis, restoring ischemia damage and promoting cell growth and used for treating skin defects, skin ulcers, pressure sores, bone defects, bone ununion, femoral head necrosis, renal injury, ischemic injury, spinal cord injury, islet damage, diabetes, complications of diabetes, Alzheimer's diseases and the like.

Description

technical field [0001] The present invention relates to biomedicine, in particular to a method and application of exosomes derived from human urine cells. Background technique [0002] Recent studies have shown that cell transplantation therapy has significant effects and broad application prospects in repairing various tissue and organ injuries. However, there are still many problems in the direct transplantation of cells for the treatment of human diseases, such as safety, immune rejection, hidden dangers of tumor formation, and difficulties in transportation and storage. Recent studies have suggested that cells perform their function of repairing tissue damage mainly through a paracrine mechanism, and exosomes secreted by cells in the paracrine mechanism may be a key component for cells to function. [0003] Exosomes are multivesicular bodies of eukaryotic cells and cell membranes, which are secreted to the outside of the double-membrane-wrapped structure after fusion. I...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61F13/02A61K35/22A61P17/02A61P19/08A61P13/12A61P25/00A61P1/18A61P3/10A61P25/28
CPCA61F13/02A61K35/22C12N5/0687
Inventor 汪泱牛鑫张长青李青胡斌姜珍珍
Owner 上海艾棵颂生物科技有限公司
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