The application of type 1 thioredoxin in the culture of endothelial progenitor cells and the culture method of endothelial progenitor cells

Abstract

The invention relates to the field of cell culture, and particularly relates to application of thioredoxin in culture of endothelial progenitor cells (EPCs) as well as a culture method of the EPCs. The invention provides the application of the thioredoxin in delaying senility of the EPCs and improving the multiplication efficiency of the EPCs, and also provides a culture solution and a culture method for culturing the EPCs. According to the application and method, a plurality of growth factors are added into a basal medium and the thioredoxin is also added, so as to well promote the growth of the EPCs. The good growth activity and stem cell property of the EPCs are maintained. The experiment provided by the invention confirms that by adding the thioredoxin into the culture solution for culturing the EPCs, the 7th generation of cell passage can still keep vigorous growth and good activity, at the 10th generation of the cell passage, the expression quantity of the surface marker CD309 is 98 percent, and the stem cell property keeps well.

Description

Technical field [0001] The invention relates to the field of cell culture, in particular to the application of type I thioredoxin in the culture of endothelial progenitor cells and a method for culturing endothelial progenitor cells. Background technique [0002] At present, the endothelial seed cells used to construct tissue-engineered blood vessels are mostly mature endothelial cells digested from the large blood vessels of adult animals or humans. This not only causes new body trauma, but also the endothelial cells from this source expand in the process of in vitro culture. The amount and cell activity are not ideal, and it is difficult to achieve the desired effect. Therefore, a vascular endothelial cell with excellent proliferation ability and activity is urgently needed as seed cell. [0003] Endothelial progenitor cells (EPCs), the precursor cells of vascular endothelial cells, are a subset of CD34-positive cells. In 1997, it was reported in the literature that CD34-positi...

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Problems solved by technology

[0004] However, most of the existing technologies are not ideal for the cultivation of EPCs, and there are some common problems that lead to the inability of EPCs to proliferate stably in vitro, such as: limited number of days of proliferation (usually within 12 days), limited number of passages (usually within P6 generation) ), the multiplication factor is not high (usually below 50 times), and it is easy to age (the longer the cell culture time, the lower the expression of cell surface markers CD34+, VEGFR-2 (also written as CD309)+); Large, the number of endothelial progenitor cells cultured from different human samples is quite different, and cannot be produced stably; the above-mentioned problems have led to the inability to produce stable EPCs, which in turn leads to the inability to use EPCs well in clinical practice

[0005] The problem leading to the poor proliferation of EPCs is mainly the formula of the culture medium. In the traditional formula, fetal bovine serum is usually added to maintain the activity of EPCs. However, the addition of heterologous serum increases the risk of human body rejection. Moreover, heterologous Although serum is rich in nutrients, its effect on maintaining cell activity is actually very limited, and it cannot meet the clinical needs for the effect of EPCs culture

[0006] Thioredoxin is a kind of small acidic protein with redox activity in cells, which has anti-oxidation and promotes angiogenesis, but its activity in vitro has not been confirmed

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