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Endothelial cell bioreactor as well as preparation method and application thereof

A bioreactor, endothelial cell technology, applied in biochemical equipment and methods, microorganisms, biochemical instruments, etc., can solve the problems of poor growth and high requirements for endothelial cell growth environment

Inactive Publication Date: 2012-07-04
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Endothelial cells have high requirements on the growth environment and cannot grow well in ordinary hollow fiber bioreactors
[0006] Endothelial cells play an extremely important role in the occurrence and development of sepsis. At present, there are no reports about endothelial cell bioreactors and their application in severe sepsis at home and abroad.

Method used

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  • Endothelial cell bioreactor as well as preparation method and application thereof
  • Endothelial cell bioreactor as well as preparation method and application thereof
  • Endothelial cell bioreactor as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Endothelial Cell Culture

[0058] Porcine iliac artery endothelial cell line (purchased from the Cell Bank of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences). This experiment was completed in the 3rd to 4th generation of this cell line. Pig iliac artery endothelial cells were grown in RPMI1640 medium containing 10% fetal bovine serum, double antibodies (0.0606g / L penicillin and 0.1g / L streptomycin), 2mmol / L L-glutamine, at 5% CO2, 37 ° C incubator incubation. When the PIEC cells normally cultured in the 75cm2 culture flask reached 90% confluency, they were washed once with PBS and digested by adding 0.25% trypsin-0.02% EDTA 1:1 mixture. Add 5ml of culture medium containing serum and double antibody, gently pipette to form cell suspension, and subculture.

Embodiment 2

[0059] Example 2 Preparation of non-woven endothelial cell bioreactor

[0060] 1. Pipeline connection of the culture fluid circulation artificial capillary culture system The culture fluid circulation artificial capillary culture system is mainly composed of 4 parts: reactor, circulation pipeline, oxygenator and peristaltic pump. The specific connection method of the pipeline is as follows: Figure 5 shown. The oxygenator is equipped with 250ml RPMI 1640+10% FBS culture solution, and communicates with the outside air through a filter. Before the pipeline is connected, the inner cavity and the outer cavity of the reactor are pre-washed with physiological saline, and the outer cavity is closed after the physiological saline is emptied, and the inner cavity is connected with the circulation pipeline. After the tubing is connected, start the peristaltic pump to evacuate the gas in the inner cavity.

[0061] 2. Digestion and counting of endothelial cells when 75cm 2 After the PI...

Embodiment 3

[0066] Embodiment 3 animal experiments

[0067] 1. Establishment of porcine endotoxemia model

[0068] Ten healthy hybrid domestic pigs (purchased from the Animal Experiment Center of Fudan University School of Medicine), male or female, about 3 months old, weighing 15kg-20kg, were divided into endothelial cell bioreactor group (experimental group) and pseudocirculation group ( control group) each with 5 heads. The experimental pig was anesthetized with 15 mg / kg of ketamine, and a venous cannula was placed in the ear vein, and the balance fluid was given at a rate of 20ml / kg / h, and 10ml / kg / h was maintained after 1 hour, and then the trachea was intubated and treated with isoflurane. Intravenous inhalation compound anesthesia was maintained, and then a double-lumen catheter was inserted into the internal carotid artery, and a floating catheter was placed. After stabilization, intravenous infusion of endotoxin (LPS) at a dose of 0.25 mg.kg-1 was completed within 0.5 hours, fol...

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Abstract

The invention belongs to the field of organizational project and medical materials and relates to an endothelial cell bioreactor as well as a preparation method and an application thereof. A polyfluortetraethylene non-woven fabric is used as a carrier for the growth of endothelial cells, and the polyfluortetraethylene non-woven fabric and a hollow fiber bioreactor constitute the bioreactor. The non-woven fabric is positioned in a clearance outside the hollow fiber so as to form a reactor which takes the polyfluortetraethylene non-woven fabric as the carrier. The endothelial cells are grown on the non-woven fabric so as to form the endothelial cell bioreactor. The number of the endothelial cells in the bioreactor is more than 3*10<9>. The endothelial cell bioreactor can improve hemodynamical indexes of organisms suffering sepsis and the disorder state of inflammation and hemoglutination, prolong the life time and improve the prognosis. The endothelial cell bioreactor can be used for curing severe sepsis.

Description

technical field [0001] The invention belongs to the field of tissue engineering and medical materials, in particular to an endothelial cell bioreactor, its preparation method and its application in treating severe sepsis. Background technique [0002] Sepsis is a systemic inflammatory response syndrome caused by infection, which is characterized by high morbidity and mortality and poor treatment effect. Sepsis has been a serious threat to human life. Epidemiological surveys show that there are more than 18 million severe sepsis patients worldwide every year, and the number of patients is increasing at a rate of 1.5% per year. In the United States, the mortality rate of sepsis is 28.6%, the mortality rate of severe sepsis is as high as 30%-50%, and the mortality rate of septic shock is 40%-60%. The mortality rate of sepsis has exceeded that of acute myocardial infarction. infarction. Therefore, in October 2002, many organizations in European and American countries jointly ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/00C12N5/06A61F2/00A61P31/00
Inventor 丁峰顾勇谢琼虹刘骏峰长谷川冰李一张家瑛林善锬平易梁伟民朱建国
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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