Method for producing epidermal growth factors in mixed cell and simulation cell culture artificial niche device

Abstract

The invention relates to a method for producing epidermal growth factors in a mixed cell and simulation cell culture artificial niche device, and belongs to the field of intelligent manufacturing of cytokine biological products. The method for producing epidermal growth factors comprises the steps of firstly, preparing mixed cell culture liquid and cell nutrition liquid, then setting parameters ofthe artificial niche device and loading the mixed cell culture liquid in stem cell niches, starting a circulating device for cell culture, and finally, performing cytokine detection on collected cellsupernatant by using an ELISA reagent kit. The artificial niches are used for establishing temperature, nutrient, acid-base balance, oxygen balance, carbon dioxide balance and metabolites discharge simulation conditions in human bodies when the epidermal growth factors are produced in human bodies, so that the synthesis of the epidermal growth factors is closer to the condition of the human bodycompared with that under a conventional 2D cell culture condition.

Description

technical field [0001] The invention belongs to the field of intelligent manufacturing of cytokine biological products, and in particular relates to a method for producing epidermal growth factor in a mixed cell and simulated cell culture artificial nest device. Background technique [0002] Cytokines are a class of biologically active small molecular proteins secreted by cells. Cytokines generally regulate cell growth, differentiation, and effects by binding to corresponding receptors. Cytokines serve as molecular messengers that allow immune system cells to communicate with each other to produce anti-target The coordination of antigens, regulatory and effector functions in many diseases, so cytokines and their receptors can be used in immunotherapy. [0003] Due to the high efficiency of cytokines, cytokines are clinically used in the treatment of various diseases such as tumors and metabolic diseases, but there are many problems. Most of the time, the occurrence and devel...

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Problems solved by technology

[0003] Due to the high efficiency of cytokines, cytokines are clinically used in the treatment of various diseases such as tumors and metabolic diseases, but there are many problems. Most of the time, the occurrence and development of diseases are not determined by a single type of cell and a single physical and chemical factor The impact of cytokines on diseases depends more on the homeostasis and balance of the cytokine network composed of multiple cytokines. Most of the clinical applications of cytokines are not endogenous cytokines produced by patients themselves. Many cells and molecular network relationships in the body are still unknown, which leads to the fact that the clinical treatment effect of many cytokines is far inferior to the in vitro effect; secondly, single cytokine therapy can easily lead to severe cytokine toxicity in the body, and the cytokines Short half-life, easy to lose biological activity

[0004] One of the important methods of traditional epidermal factor production is to use gene recombination technology to construct transgenic cells to produce epidermal growth factor under 2D cell culture conditions. Viral vectors can continuously and effectively express the target gene, but there are still doubts about the expression and safety of viral proteins; adenoviral vectors will not fuse with host genes, sometimes require repeated transduction, and may trigger immune and inflammatory responses; adeno-associated viral vectors It has the characteristics of low harmfulness, wide application range, and stable target gene product, but it is still being improved due to the problem of low utilization efficiency

Commonly used non-viral vectors include liposomes, polymers, etc. Liposomes are likely to cause immune recognition reactions and be degraded by the reticuloendothelial system due to the complex macromolecules formed by liposomes and target cells. Potential toxicity and efficiency of polymer carriers

Secondly, in order to pursue the difference between in vivo synthesis and in vitro synthesis, the production methods of many protein molecules adopt the method of using mammalian cells to produce protein molecules. However, the culture of mammalian cells often relies on serum culture, but this Traditional culture methods can easily cause mycoplasma and other virus contamination

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