System for producing and preparing exosome and method for preparing exosome

An exosome and system technology, applied in the field of biomedicine, can solve the problems of time-consuming and labor-intensive, low recovery rate, and inability to industrialize large-scale production of exosomes.

Inactive Publication Date: 2021-09-14
瑞太生物科技(沈阳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] For this reason, the present invention provides a system for the production and preparation of exosomes and a method for preparing exosomes to solve the problem of time-consuming and labor-consuming extraction of exosomes, low recovery rate, serious pollution in the intermediate process, and low yield. , unable to industrialize large-scale production and other problems

Method used

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  • System for producing and preparing exosome and method for preparing exosome
  • System for producing and preparing exosome and method for preparing exosome
  • System for producing and preparing exosome and method for preparing exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 The present invention provides a method for preparing exosomes using the exosome preparation system

[0036] figure 1 Preparation of system diagrams for large-scale production of exosomes; figure 2 It is the structural diagram of the hollow fiber cell culture device.

[0037] 1. Inoculation and culture of cells

[0038] Add the culture medium into the first storage tank 1, turn on the first peristaltic pump 2, pump the culture in the first storage tank 1 into the hollow fiber cell culture device 3, and fill it up, take 10 7 Umbilical cord mesenchymal stem cells cultured to the third generation are inoculated into the hollow fiber cell culture device 3 through the cell inoculation port 7, shaken evenly, and cultured at 25°C for 10 hours, so that the umbilical cord mesenchymal stem cells are fully attached to the hollow fiber tube surface.

[0039] 2. Large-scale production and preparation system of exosomes to extract exosomes

[0040]Observe the culture ...

Embodiment 2

[0043] Example 2 A system for the production and preparation of exosomes

[0044] The system for preparing exosomes comprises a first storage tank 1, a second storage tank 4, a hollow fiber cell culture device 3, a tangential flow filtration device 6, a first peristaltic pump 2, and a second peristaltic pump 5;

[0045] Wherein, the outlet of the first storage tank 1 is connected with the inlet of the hollow fiber cell culture device 3 through the first peristaltic pump 2, the outlet of the hollow fiber cell culture device 3 is connected with the inlet of the first storage tank 1, and the filtrate of the hollow fiber tube 3 The outlet is connected with the first inlet of the second storage tank 4; the outlet of the second storage tank 4 is connected with the inlet of the tangential flow filter device 6 through the second peristaltic pump 5, and the outlet of the tangential flow filter device 6 is connected with the second storage tank 4 is connected to the second inlet, and th...

experiment example 1

[0050] Experimental example 1 Identification of exosomes and comparison of biological activities of exosomes extracted by different methods

[0051] 1. Electron microscope comparison of exosomes

[0052] Using glutaraldehyde solution, the exosomes obtained in Example 1 and Comparative Example were respectively fixed on the sample-loading copper grid, and after reacting for 5 minutes, the 2 O wash the copper grid, then place the copper grid in uranyl oxalate solution and incubate for 5 minutes, and finally absorb the excess liquid with filter paper. After the copper grid is dry, put it in the sample box, and take electron microscope photos at 80kV. like Figure 5 As shown in the electron microscope examination image of the exosomes extracted and purified in the example, the vesicle structure of the saucer-shaped double-layer membrane can be seen under the electron microscope, the particle size is in the range of 30-150nm, and the background is clean, the structure is clear, an...

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Abstract

The invention discloses a system for producing and preparing exosome and a method for preparing the exosome, and particularly relates to the field of biological medicine. The method comprises two parts of cell culture and exosome extraction. The system comprises a first storage tank, a second storage tank, a hollow fiber cell culture device, a tangential flow filtering device, a first peristaltic pump and a second peristaltic pump. The system can be applied to industrial large-scale production and preparation of the exosome. According to the system, a large number of hollow fiber tubes in the hollow fiber device are utilized, so that the cell culture area is increased; cells can be promoted to secrete a large amount of exosome through a culture medium conveyed by the hollow fiber tubes, and the produced exosome solution is purified through the tangential flow filtering device; the system is simple to operate, high in recovery rate, free of intermediate pollution, high in purity and high in concentration. By means of the system, the integrated preparation process from cell culture to exosome production can be realized, and the system is very suitable for industrial large-scale production and preparation of clinical-grade exosome.

Description

technical field [0001] The present invention relates to the technical field of biological medicine, in particular to a system for producing and preparing exosomes and a method for preparing exosomes. Background technique [0002] Exosomes are small vesicles with a phospholipid bilayer structure secreted by living cells, with a diameter of 30-150nm and a density of 1.13-1.19g / ml, which can exist in various body fluids, such as serum, plasma, saliva, urine fluid, ascites, spinal fluid, milk, etc. Exosomes contain a variety of biomolecules, such as mRNA, miRNA, proteins, lipids, etc., which can be delivered to recipient cells, thereby changing the physiological or pathological functions of recipient cells. In recent years, exosomes have attracted widespread attention as intercellular information transfer tools and biomarkers of various diseases. Exosomes have great potential in the fields of biomedicine and disease diagnosis. [0003] Obtaining high-purity, high-yield, and st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/12C12N5/00C12N5/0775
CPCC12M25/10C12M47/12C12N5/00C12N5/0668
Inventor 张海心
Owner 瑞太生物科技(沈阳)有限公司
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