Application of umbilical cord mesenchymal stem cells in the preparation of pharmaceutical preparations for the treatment of pulmonary interstitial fibrosis

A technology for pulmonary interstitial fibrosis and stem cells, which can be applied to animal cells, drug combinations, vertebrate cells, etc., can solve the problem of not obtaining positive results in the bleomycin lung injury model, and achieve low preparation cost and no treatment. Side effects, the effect of inhibiting the deposition of collagen fibers

Active Publication Date: 2019-08-09
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT) +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Nevertheless, some studies using the bleomycin lung injury model have not yielded positive results

Method used

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  • Application of umbilical cord mesenchymal stem cells in the preparation of pharmaceutical preparations for the treatment of pulmonary interstitial fibrosis
  • Application of umbilical cord mesenchymal stem cells in the preparation of pharmaceutical preparations for the treatment of pulmonary interstitial fibrosis
  • Application of umbilical cord mesenchymal stem cells in the preparation of pharmaceutical preparations for the treatment of pulmonary interstitial fibrosis

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Isolation, culture and identification of umbilical cord mesenchymal stem cells

[0035] 1 Isolation and culture of umbilical cord mesenchymal stem cells

[0036] The umbilical cord ligated after delivery of the fetus was taken, washed and disinfected, placed in umbilical cord preservation solution, and kept at a constant temperature of 6°C. Cut the Wharton's colloid to 2 mm with sterile tissue scissors 3 Add appropriate amount of medium to the tissue homogenate block, and place it in a carbon dioxide constant temperature and humidity incubator for culture. On the 16th day, when the area percentage of the cell clone reached 80%, it was digested and harvested to obtain P0 generation cells. Afterwards, digestion and passage were carried out, and the cells to be frozen were placed in a programmed cooling device, and the cells were lowered to below -80°C according to the standard freezing procedure.

[0037] 2 Identification of surface antigens of umbilical cord mesenchymal...

Embodiment 2

[0041] Effects of IFN-γ treatment on phenotype and secretion of cytokines and chemokines of umbilical cord mesenchymal stem cells

[0042] The P4 generation umbilical cord mesenchymal stem cells were mixed at 1×10 per well 5 Planted in a 6-well plate, changed to serum-free medium after 24 hours to starve the cells for 12 hours, added 20ng / ml recombinant human IFN-γ for 24 hours, collected the supernatant, and detected the anti-fibrosis effect by real-time fluorescent quantitative PCR The mRNA expression changes of cytokines and chemokines (IP-10, PGE2), the experimental results are shown in figure 1 , the cells were collected, and the changes of cell surface immune markers (CD90, CD29, CD73, CD105, CD45, CD34, CD19, CD14, HLA-DR) were detected by flow cytometry. The experimental results are shown in Table 1.

[0043] Table 1 The immunophenotype results of umbilical cord mesenchymal stem cells before and after IFN-γ pretreatment

[0044] cell surface immunolabeling ...

Embodiment 3

[0049] Preparation of culture supernatant of umbilical cord mesenchymal stem cells pretreated with IFN-γ and expression of cytokines, chemokines IP-10 and PGE2

[0050] The mesenchymal stem cells were treated with 20ng / ml recombinant human IFN-γ for 24 hours, washed 3 times with PBS, added serum-free medium to continue culturing for 48 hours, and the culture supernatant was collected to be the supernatant required for subsequent animal experiments. The expression of IP-10 and PGE2 was detected by ELISA method, the experimental results are shown in figure 2 .

[0051] It was found that, if figure 2 As shown, in the culture supernatant of umbilical cord mesenchymal stem cells pretreated with IFN-γ, the levels of IP-10 and PGE2 were significantly higher than those of the untreated group.

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Abstract

The invention discloses an application of umbilical cord mesenchymal stem cells in preparation of a pharmaceutical preparation for treating the PF (pulmonary fibrosis). When FN-gamma pretreated mesenchymal stem cells are adopted to treat PF of mice, the survival rate of the mice can be increased obviously, BLM (bleomycin) induced PF pulmonary lesion can be relieved, the pulmonary histopathologic score of the mice can be reduced obviously, and the umbilical cord mesenchymal stem cells have broad application prospect in preparation of medicines for treating the PF.

Description

technical field [0001] The present invention relates to the use of mesenchymal stem cells, in particular to the application of the culture supernatant of umbilical cord mesenchymal stem cells in the treatment of pulmonary interstitial fibrosis. Background technique [0002] Pulmonary fibrosis (Pulmonary Fibrosis, PF) is the abbreviation of diffuse pulmonary fibrosis. disease. The patient's condition is serious, and the case fatality rate is high, seriously endangering human health. So far, its etiology and pathogenesis are still unclear, and there is still no effective treatment in clinical practice. Although hormones have a certain effect on non-specific interstitial pneumonia, which is dominated by inflammation, they cannot slow down the process of pulmonary fibrosis and lung function decline, so the therapeutic effect on idiopathic pulmonary fibrosis (Idiopathic Pulmonary Fibrosis, IPF) is poor. Lung transplantation is one of the optional treatment strategies, but limi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61K35/28A61K35/51A61P11/00
Inventor 刘明徐军
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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