Application of MSC (mesenchymal stem cell) exosomes in preparation of pharmaceutic preparation for treating PF (pulmonary fibrosis)

A technology of mesenchymal stem cells and pharmaceutical preparations, applied in the application field of mesenchymal stem cell exosomes in the treatment of pulmonary fibrosis, can solve the problems of non-expression and no detection of MSC-specific surface markers, and achieve simple methods and low preparation costs Inexpensive, the effect of improving the survival rate

Active Publication Date: 2015-06-03
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specific surface markers of MSC have not been found so far, and the uniform and stable antigen markers of MSC include: SH2(+), SH3(+), CD29(+), CD44(+), CD71(+), CD90(+), CD106( +), CD120a(+), CD124(+), does not express hematopoietic cell surface markers CD34, CD45, CD14

Method used

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  • Application of MSC (mesenchymal stem cell) exosomes in preparation of pharmaceutic preparation for treating PF (pulmonary fibrosis)
  • Application of MSC (mesenchymal stem cell) exosomes in preparation of pharmaceutic preparation for treating PF (pulmonary fibrosis)
  • Application of MSC (mesenchymal stem cell) exosomes in preparation of pharmaceutic preparation for treating PF (pulmonary fibrosis)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Isolation and identification of exosomes from mesenchymal stem cells and normal lung fibroblasts

[0034] 1 Isolation, culture and identification of mesenchymal stem cells and normal lung fibroblasts (NLF)

[0035] Under sterile conditions, cut the umbilical cord into small pieces with a length of about 1 cm, wash them thoroughly, remove the arteries and veins of the umbilical cord, cut the small pieces of the umbilical cord into small pieces, place them in a petri dish, use Wharton glue to directly adhere to the bottom of the dish, and add a small amount of 10wt% FBS and 5wt% green chain double antibody low-sugar DMEM culture solution, placed at 37 ° C, 5% CO 2 Cultivate in an incubator, and add culture medium after 4 hours. After 3 days, the culture medium was changed, and the medium was changed every 3 days. After the MSCs reached 70-80% fusion, they were digested with 0.25% trypsin, subcultured, and the P3 generation cells were taken for experiments.

[0036] Tak...

Embodiment 2

[0043] MSC exosomes can enter target cells and inhibit the expression of α-SMA and Fibronectin in abnormally activated fibroblasts

[0044] 1MSC exosomes can enter target cells (human lung fibroblasts)

[0045] The extracted MSC exosomes and normal lung fibroblast exosomes were labeled with CM-Dil from Invitrogen, and added to the adherent cultured normal human lung fibroblasts. After 24 hours, the exosomes entered with a fluorescence microscope were observed. In the case of cells, the results are as follows image 3 As shown, both red fluorescent-labeled MSC exosomes and lung fibroblast exosomes can successfully enter human lung fibroblasts.

[0046] 2MSC exosomes can inhibit the expression of α-SMA and Fibronectin in normal human lung fibroblasts activated by TGF-β1

[0047] Isolate and culture normal human lung fibroblasts, plant P3 generation cells in a 6-well plate at an appropriate density, and starve the cells for 12 hours after the cells have reached 70% confluence. ...

Embodiment 3

[0053] Intratracheal administration of MSC exosomes reduces mortality and lung lesions in BLM-induced pulmonary fibrosis

[0054] 1 Establishment of BLM-induced pulmonary fibrosis mouse model

[0055] Take 6-week-old SPF grade BALB / C mice and anesthetize them by intraperitoneal injection of pentobarbital sodium. Under the direct vision of the laryngoscope, use the airway of small animals to administer drugs, and insert the drug catheter into the trachea with a high-pressure gun. The dose of kg was injected into PBS, and the experimental control group and the two treatment groups were injected into BLM at a dose of 3 mg / kg. After the injection, the animal was upright and rotated to make the drug evenly distributed in the lungs.

[0056] 2 Intratracheal administration of MSC exosomes reduces survival in BLM-induced pulmonary fibrosis

[0057] On the second day after BLM administration, the MSC exosome treatment group was given 50ul of MSC exosomes at a concentration of 1ug / ml i...

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Abstract

The invention discloses an application of MSC (mesenchymal stem cell) exosomes in preparation of a pharmaceutic preparation for treating PF (pulmonary fibrosis). The MSC exosomes are adopted to treat PF of a mouse, the survival rate of the mouse can be remarkably increased, the PF pulmonary lesions caused by BLM (bleomycin) are relieved, the mouse lung tissue pathological score is reduced, and accordingly, the MSC exosomes have wide application prospect in the aspect of preparation of the pharmaceutic preparation for treating PF.

Description

technical field [0001] The present invention relates to the use of mesenchymal stem cell exosomes, in particular to the application of mesenchymal stem cell exosomes in the treatment of pulmonary fibrosis. Background technique [0002] Pulmonary fibrosis (PF), also known as diffuse pulmonary parenchymal disease, is a diffuse lung disease involving pulmonary interstitium, alveoli, and bronchioles caused by various reasons, which seriously endangers human health. Its etiology is complicated, and the clinical treatment effect is poor. Idiopathic pulmonary interstitial fibrosis and nonspecific interstitial pneumonia are the two most common types, especially idiopathic pulmonary interstitial fibrosis is barely responsive to current treatments including glucocorticoids and cytotoxic drugs , poor prognosis, the average 5-year survival rate is only 30 to 50%. Pulmonary fibrosis usually occurs between the ages of 40 and 50, and it occurs more frequently in men than in women. Dyspn...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K9/00A61P11/00C12N5/0775
Inventor 刘明徐军
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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