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Application of exosome in promotion of growth of mesenchymal stem cells of decidua parietalis

A technology of mesenchymal stem cells and exosomes, which is applied to promote the growth of decidual mesenchymal stem cells. The application field of exosomes in promoting the growth of decidual mesenchymal stem cells can solve the problem of easy aging of PDB-MSCs and the appearance of exosomes. The content and source of secretions are limited, and it is difficult to reproduce the experimental results, etc., to achieve the effect of good cell shape, fast and simple extraction method, and strong cell proliferation ability

Pending Publication Date: 2021-07-23
GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, at present, the application of PDB-MSCs still faces some problems: the existing methods and technologies can isolate PDB-MSCs with high purity, but after passage, PDB-MSCs are prone to aging, flat cells, slow or stop proliferation, and loss of differentiation ability etc. Therefore, new methods or technologies are needed to effectively maintain the morphology of PDB-MSCs, prevent cell aging, and maintain cell proliferation and differentiation after multiple passages.
[0005] The patent with the publication number CN105349487 discloses a method for promoting the proliferation of human bone marrow mesenchymal stem cells based on exosomes, using exosomes secreted by first-generation or second-generation human umbilical cord mesenchymal stem cells to promote the proliferation of human bone marrow. The proliferation of mesenchymal stem cells, the content and source of exosomes are greatly restricted
In addition, the exosome-promoting medium for human bone marrow mesenchymal stem cells contains 5% UltraGRO by mass percent and α-MEM basal medium containing 0.032% medical heparin sodium by mass percent.
Secondly, when the experiment of promoting the proliferation of mesenchymal stem cells is scaled up, the operation is more cumbersome.

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  • Application of exosome in promotion of growth of mesenchymal stem cells of decidua parietalis
  • Application of exosome in promotion of growth of mesenchymal stem cells of decidua parietalis
  • Application of exosome in promotion of growth of mesenchymal stem cells of decidua parietalis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A method for preparing human umbilical cord mesenchymal stem cell exosomes, comprising the following steps:

[0044]S11. The hospital collects the placental tissue of the full-term caesarean section fetus, signs the customer's informed consent before the collection, and transports the tissue to the laboratory in a refrigerated and sterile environment at 4°C. During the transportation, the tissue protection solution is used to protect the biological activity of the placental tissue and ensure the transportation process. There is no bacterial and fungal contamination in the medium; the tissue protection solution is prepared from 1 ml of normal saline, 25 μg of gentamicin sulfate, and 5 μg of amphotericin B.

[0045] S12. In the laboratory, use tissue protection solution to clean the blood on the surface of the umbilical cord tissue, remove the epidermis and vascular tissue, take out Huatong glue, and cut it into 1-2mm pieces after cleaning 3 Fragments, inoculated in T75 c...

Embodiment 2

[0053] A method for promoting the proliferation of parietal decidual mesenchymal stem cells, comprising the following steps:

[0054] S21. Prepare cell culture media containing exosomes at different concentrations. The human umbilical cord mesenchymal stem cell exosomes prepared in Example 1 are prepared at different concentrations (0 μg / ml, 10 μg / ml, 20 μg / ml and 30 μg / ml). ml) were added to different serum-free medium for mesenchymal stem cells (Youkang Hengye Biotechnology (Beijing) Co., Ltd., product number: NC0103);

[0055] S22, P12 generation wall decidual mesenchymal stem cells according to 2 × 10 per well 3 The concentration of cells is inoculated in a 96-well plate, and the cell culture medium containing exosomes in step S21 is added respectively;

[0056] S13, placed at 37°C, 5% CO 2 Cultivated in a saturated humidity carbon dioxide incubator.

[0057] Each set of experiments was repeated at least three times.

[0058] PDB-MSCs were collected and counted every 2...

Embodiment 3

[0061] A method for promoting the proliferation of parietal decidual mesenchymal stem cells, comprising the following steps:

[0062] S21. Prepare cell culture media containing exosomes at different concentrations. The human umbilical cord mesenchymal stem cell exosomes prepared in Example 1 are prepared at different concentrations (0 μg / ml, 10 μg / ml, 20 μg / ml and 30 μg / ml). ml) were added to different serum-free medium for mesenchymal stem cells (Youkang Hengye Biotechnology (Beijing) Co., Ltd., product number: NC0103);

[0063] S22, P12 generation wall decidual mesenchymal stem cells according to 4 × 10 per well 5 The concentration of cells is inoculated in a 96-well plate, and the cell culture medium containing exosomes in step S21 is added respectively;

[0064] S13, placed at 37°C, 5% CO 2 Cultivated in a saturated humidity carbon dioxide incubator.

[0065] Each set of experiments was repeated at least three times.

[0066] PDB-MSCs were collected after 48 hours of c...

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Abstract

The invention discloses application of a human umbilical mesenchymal stem cell exosome in promotion of growth of mesenchymal stem cells of decidua parietalis and a method for promoting the growth of the mesenchymal stem cells of the decidua parietalis. According to the application and the method, after the human umbilical mesenchymal stem cell exosome and PDB-MSCs are cultured jointly, the PDB-MSCs is high in cell proliferation capability, good in cellular form and high in cellular activity, thus, the human umbilical mesenchymal stem cell exosome disclosed by the invention can be used for improving the quality of the PDB-MSCs, the cell factor VEGF and SCF secreting capability of the PDB-MSCs is improved effectively, thus, the problems of the PDB-MSCs that repeated transfer of generation is lowered in proliferation capability and poor in cell activity can be solved, and then, the large-scale culture and clinical application of the PDB-MSCs is facilitated effectively.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an application of exosomes in promoting the growth of decidual mesenchymal stem cells and a method for promoting the growth of decidual mesenchymal stem cells. Background technique [0002] Parietal decidual basalis-mesenchymal stem cells (PDB-MSCs) have the common characteristics of mesenchymal stem cells such as multilineage differentiation potential and low immunogenicity. Compared with other sources of mesenchymal stem cells, PDB-MSCs are derived from the mother, which has better compatibility with the mother, more uniform purity, and stronger adipogenic differentiation ability than mesenchymal stem cells from other perinatal tissues; It has also been reported that PDB-MSCs secrete more vascular endothelial growth factor (VEGF) and stem cell growth factor (SCF) than umbilical cord mesenchymal stem cells, indicating that PDB-MSCs can promote angiogenesis and cell division. MSCs p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2502/137C12N2509/00C12N2509/10C12N2500/90C12N5/0682C12N2502/1388C12N2500/99C12N5/0081C12N2500/02C12N2502/025C12N2500/84
Inventor 刘小翠蒙燕瑶唐淑艳许峻荣雅思敏杨景利王进辉
Owner GUANGDONG VITALIFE BIOTECHNOLOGY CO LTD
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