Application of mesenchymal stem cell-derived exosome in preparing drug or preparation for treating preeclampsia

A kind of stem cell, preeclampsia technology, applied in the application field of exosomes in the treatment of preeclampsia, to achieve the effect of alleviating symptoms, reducing inflammatory response, and a wide range of sources

Active Publication Date: 2016-07-20
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there has been no report on the treatment of PE with exosomes derived from MSCs. For the first time, we applied the exosomes produced by MSCs derived from human umbilical cord to the treatment of PE, and achieved a good effect similar to that of MSCs.

Method used

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  • Application of mesenchymal stem cell-derived exosome in preparing drug or preparation for treating preeclampsia
  • Application of mesenchymal stem cell-derived exosome in preparing drug or preparation for treating preeclampsia
  • Application of mesenchymal stem cell-derived exosome in preparing drug or preparation for treating preeclampsia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Isolation and culture of human umbilical cord mesenchymal stem cells (MSCs) (the reagents and consumables used in the laboratory are all aseptically processed)

[0023] With the informed consent of the puerpera, the umbilical cords of the cesarean section fetuses of normal full-term pregnancy were collected. Before the umbilical cord is collected, the puerpera needs to undergo strict pathogen testing, including Treponema pallidum, HIV, cytomegalovirus, hepatitis B virus, hepatitis C virus, syphilis and mycoplasma, and use it after confirming safety.

[0024] Cut the umbilical cord near the placenta to 20-30 cm, store it in pre-cooled sterile phosphate buffered saline (PBS) at 4°C, and use it within 4 hours. In the ultra-clean workbench, the umbilical cord was cut into small sections of about 5 cm, and the residual blood on the surface of the umbilical cord was washed with PBS; DMEM / F12 medium (purchased from Gibco, the same below), cut into 2mm with surgical ...

Embodiment 2

[0026] Example 2 Extraction and Identification of MSCs-derived Exosomes

[0027] (1) Extraction of MSCs-derived exosomes

[0028] Collect the MSCs culture supernatant of passages 3-7 into 500ml sterile centrifuge bottles or 50ml polypropylene centrifuge tubes (purchased from Beckman), and centrifuge at 4°C and 2000g for 10 minutes to remove dead cells and large debris. Carefully transfer the supernatant to a new sterile centrifuge tube and centrifuge at 10,000g for 30 minutes at 4°C to remove organelles and small particles. Carefully transfer the supernatant to a sterile ultracentrifuge tube, and centrifuge at 110,000g (Beckman ultracentrifuge) at 4°C for 70 minutes, discard the supernatant carefully, wash once with normal saline for injection, and store at 4°C, 110,000g ultracentrifugation for 70 minutes, the obtained pellet is exosomes. Depending on the volume of the culture medium collected initially, a small volume of normal saline for injection was added as appropriate ...

Embodiment 3

[0033] Example 3 The therapeutic effect of MSCs and their exosomes on preeclampsia (PE) mice

[0034] The C57 / B6 mice used were 8-10 weeks old and purchased from the Institute of Model Animals, Nanjing University.

[0035] (1) Low-dose lipopolysaccharide (LPS) induced mouse PE model

[0036] Mice were housed in a female-male ratio of 2:1, and vaginal plugs were checked in the morning. Those with vaginal plugs were considered to have successfully conceived, recorded as the 0.5th day of pregnancy, and so on. Pregnant mice were randomly divided into 4 groups: control group, model group, MSC treatment group and MSC-derived exosome treatment group, with 8 mice in each group. Referring to the method of Tiziana et al. (TizianaCotechinietal.Inflammationinratpregnancyinhibitsspiralarteryremodelingleadingtofetalgrowthrestrictionandfeaturesofpreeclampsia.JExpMed.2014,211(1):165-179), low-dose LPS was used to induce PE-like symptoms in mice. Model group: From day 13.5 to day 16.5, intra...

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Abstract

The invention discloses application of umbilical cord mesenchymal stem cell-derived exosome in preparing drug or preparation for treating preeclampsia, in particular relates to application of treating preeclampsia by extracting exosome through umbilical cord mesenchymal stem cells of humans. The exosome can effectively alleviate hypertension and proteinuria of preeclampsia and glomerulonephritis, reduce inflammatory response in placenta, and promote fetal development. This kind of exosome has the advantages of convenience in storage and transport and provides a new strategy for treating related diseases of refractory inflammations of preeclampsia.

Description

technical field [0001] The present invention relates to the use of exosomes derived from mesenchymal stem cells, in particular to the application of exosomes produced by mesenchymal stem cells in the treatment of preeclampsia. Background technique [0002] Preeclampsia (PE) or preeclampsia is a unique complication of pregnancy, characterized by hypertension (systolic blood pressure ≥ 140 mmHg or diastolic blood pressure ≥ 90 mmHg) and proteinuria ( ≥300mg / 24 hours or random proteinuria ≥++), which can affect various organs and systems of the body. The incidence of PE accounts for 3% to 10% of all pregnancies, and it is one of the important causes of maternal and perinatal death. Its etiology has not yet been clarified, and there is still a lack of effective prevention and treatment methods in clinical practice. [0003] Studies in recent years have shown that PE is an excessive inflammatory response of the mother to pregnancy, and low-dose lipopolysaccharide (LPS) can induc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/51A61P15/00A61P9/12C12N5/0775
CPCA61K9/0019A61K35/51C12N5/0668C12N2509/00
Inventor 侯亚义宋玉仙泥艳红樊竑冶赵晓寅赵树立
Owner NANJING UNIV
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