Preparation method of adult stem cell exosome and application thereof to treatment of Parkinson's disease

A technology of adult stem cells and exosomes, which is applied in the field of biomedicine to achieve the effect of increasing the number and restoring motor function

Pending Publication Date: 2020-07-31
YAOSHUNZE BIO MEDICAL NANJING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, there are many problems that cannot be ignored in stem cell therapy, such as bio

Method used

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  • Preparation method of adult stem cell exosome and application thereof to treatment of Parkinson's disease
  • Preparation method of adult stem cell exosome and application thereof to treatment of Parkinson's disease
  • Preparation method of adult stem cell exosome and application thereof to treatment of Parkinson's disease

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Embodiment 1

[0041]The present invention provides a method for preparing exosomes of adult stem cells, comprising: separating and culturing adult stem cells in vitro, and extracting exosomes in the supernatant of cell culture fluid by ultracentrifugation. The adult stem cells include bone marrow mesenchymal stem cells, skin epidermal stem cells and fat stem cells. Specifically include:

[0042] 1. Isolation and culture of bone marrow mesenchymal stem cells

[0043] 4-week-old male C57BL / 6J mice were anesthetized and killed by neck dislocation, and the bilateral femurs and tibias of the mice were completely extracted and rinsed with sterile normal saline and 75% ethanol solution, respectively. Remove the attached muscles and tissues on the bone surface, and use surgical scissors to cut off the bone end to expose the bone marrow cavity. Use a syringe to inhale DMEM / F-12 culture solution containing 10% fetal bovine serum, insert the needle into the bone marrow cavity, wash out the bone marr...

Embodiment 2

[0051] The adult stem cell exosomes prepared in Example 1 were administered to Parkinson's model mice by nasal administration, and behavioral testing was performed. After the behavioral testing, the mice were anesthetized and killed, and the substantia nigra was isolated for histomorphological examination. detection.

[0052] 1. Preparation of Parkinson's animal model:

[0053] Eight-week-old C57BL / 6J male mice were induced with PD model mice by intraperitoneal injection of neurotoxin MPTP (N-methyl-4-phenyl-l, 2,3,6-tetrahydropyridine). Three days before administration, the mice were subjected to a series of behavioral trainings such as rotating the rod and climbing poles every day. Try to select mice with the same level of exercise ability to avoid the influence of individual differences, and divide them into two groups. One group is the normal saline group, and the other is the MPTP group. Then intraperitoneal injection of MPTP (20mg / kg / day) was performed for one week, a...

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Abstract

The invention discloses a preparation method of an adult stem cell exosome and application thereof to treatment of Parkinson's disease, and belongs to the technical field of biomedicine. The preparation method comprises the steps that bone marrow mesenchymal stem cells, skin epidermis stem cells and adipose-derived stem cells are separated and cultured in vitro, exosomes in cell culture supernatants are extracted by an ultracentrifugation method, and the exosomes are administered by a nasal administration mode. The exosomes from the three sources can significantly relieve related symptoms of Parkinson's disease model mice, such as recovery of movement functions of the mice and increase of the number of dopaminergic neuronal cells at the black part in the brains of the mice.

Description

technical field [0001] The invention relates to a method for preparing adult stem cell exosomes and its application in treating Parkinson's disease, belonging to the technical field of biomedicine. Background technique [0002] Parkinson's disease, also known as Parkinson's disease, is the second largest neurodegenerative disease after Alzheimer's disease and seriously affects the quality of human life. According to statistics, one out of every 800 people in the world suffers from Parkinson's disease. By 2030, due to the acceleration of the aging process, the prevalence rate of Parkinson's disease will double, and it is estimated that there will be more than 9 million patients. The direct medical costs of the patients are estimated to be over US$10,000 per year, placing a heavy burden on families and society. Parkinson's disease is characterized by progressive, multiple, and insidious onset. The main manifestations are slowness of movement, muscle rigidity, resting tremor, ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61P25/16
CPCC12N5/0663C12N5/0667C12N5/0668A61P25/16C12N2509/10C12N2509/00
Inventor 顾晓松孙诚
Owner YAOSHUNZE BIO MEDICAL NANJING CO LTD
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