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Mesenchymal stem cell exosome as well as preparation method and application thereof

A technology of stem cells and exosomes, applied in the field of biomedicine, can solve the problems of long-term preservation and transportation of stem cells with abnormal differentiation and immune rejection.

Pending Publication Date: 2020-10-30
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 abnormal differentiation of stem cells, immune rejection, and long-term preservation and transportation of stem cells.

Method used

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  • Mesenchymal stem cell exosome as well as preparation method and application thereof
  • Mesenchymal stem cell exosome as well as preparation method and application thereof
  • Mesenchymal stem cell exosome as well as preparation method and application thereof

Examples

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Embodiment 1

[0027] The invention provides a method for preparing exosomes of mesenchymal stem cells, comprising: separating and culturing mesenchymal stem cells in vitro, and extracting exosomes in the supernatant of cell culture fluid by a high-speed gradient centrifugation method.

[0028] 1. Isolation and culture of bone marrow mesenchymal stem cells

[0029] Two-month-old male SD rats were killed by neck dislocation after anesthesia, and their bilateral femurs and tibias were taken, rinsed with sterile saline and 75% ethanol solution, and the two ends of the leg bones were cut off to expose the bone marrow cavity. The bone marrow cavity was washed repeatedly with DMEM / F-12 (1:100 double antibody) culture solution of %FBS, the washing solution was collected, and the cells were fully dispersed by blowing repeatedly, and then filtered through a 200-mesh stainless steel filter. Centrifuge the filtrate at 300 g for 5 min, discard the supernatant, resuspend the cells in DMEM / F-12 (1:100 dou...

Embodiment 2

[0037] The mesenchymal stem cell exosomes prepared in Example 1 were administered to the senile joint wear model rats by intra-articular administration. After 8 weeks of administration, the rats were sacrificed after anesthesia, the articular cartilage and synovial tissue were separated, and the Histology, molecular biology testing.

[0038] 1. Preparation of senile joint wear model:

[0039] The destabilization of the medial meniscus (DMM) osteoarthritis model of the knee joint can be induced by surgery, which can simulate the pathogenesis of senile bone and joint wear in a relatively short period of time. The specific method is as follows: 2-month-old male SD rats were anesthetized and fixed on the operating platform in a supine position. The skin of the rat knee joint was incised sequentially, the soft tissue was bluntly separated, the subpatellar fat tissue was removed after the patellar ligament was explored, the joint capsule was opened, and the medial meniscus and the ...

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Abstract

The invention discloses a mesenchymal stem cell exosome as well as a preparation method and an application thereof, and belongs to the technical field of biomedicine. According to the invention, bonemarrow mesenchymal stem cells, adipose mesenchymal stem cells and skin mesenchymal stem cells are separated and cultured in vitro, exosomes in a cell culture supernatant are extracted by an ultracentrifugation method, and the exosomes are administrated in an articular cavity administration mode. The three exosomes derived from mesenchymal stem cells can significantly relieve related indications ofa rat with senile knee joint wear, such as improving Mankin score of a senile knee joint wear model mouse, inhibiting inflammatory factors and activating an RANK system.

Description

technical field [0001] The invention relates to a mesenchymal stem cell exosome and a preparation method and application thereof, belonging to the technical field of biomedicine. Background technique [0002] According to the World Health Organization (WHO), approximately 355 million people worldwide suffer from osteoarthritis, a progressive and debilitating joint disease that is expected to affect up to 25% of the population by 2040. The knee joint is the largest and most complex joint in the human body, bearing 60% of the weight of the human body. Therefore, the knee joint is extremely vulnerable to wear and damage. The earliest and most important pathological changes are in the articular cartilage, which has almost no regenerative ability. After the disease occurs, with the With age, the pathological changes are irreversible and can cause knee osteoarthritis in severe cases. Most of the patients with knee osteoarthritis are elderly people. According to statistics, the in...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61K35/28A61P19/02
CPCC12N5/0663C12N5/0667C12N5/0668A61K35/28A61P19/02C12N2509/10
Inventor 孙华林顾晓松
Owner YAOSHUNZE BIO MEDICAL NANJING CO LTD
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