Efficient HaCaT cell transfection method

Abstract

The invention discloses an efficient HaCaT cell transfection method. In the process that transfection plasmids enter a HaCaT cell through a liposome method, TX-10 is added in a transfection system of the HaCaT cell. As the TX-10 is added in the transfection process, permeability of a eukaryotic cell membrane is improved, and a liposome-DNA compound is promoted to effectively enter the cell. The method is very effective for HaCaT cells thick in cell membrane and difficult to transfect through a traditional liposome method and improves the transfection efficiency by several times. The method also has the applicability for other cells thick in cell membrane and difficult to transfect; as polyethylene glycol octylphenol ether has toxicity to cells when the use amount of polyethylene glycol octylphenol ether is high and the effect is not obvious when the use amount is low, the use amount of the polyethylene glycol octylphenol ether needs to be considered. For HaCaT cells, the best dosage of the TX-10 is 0.005%; under the best dosage, the death rate of HaCaT cells is lowest and the transfection efficiency is highest.

Description

Technical field: [0001] The invention belongs to the field of biology, and in particular relates to a method for efficiently transfecting HaCaT cells. Background technique: [0002] HaCaT cells are human skin immortalized keratinocytes derived from normal human skin keratinocytes. In view of its advantages of easy culture and multiple passages, it is used as a tool cell instead of human normal keratinocytes. , through the influence of various drugs, compounds, physical and chemical factors on HaCaT cells to study the cell proliferation, cell cycle and skin pathological mechanism of human skin keratinocytes. [0003] Cell transfection refers to the process by which eukaryotic cells acquire new genetic markers due to the incorporation of foreign DNA. There are four main methods for exogenous gene transfection into cells: electric shock method, calcium phosphate method, virus-mediated method and liposome-mediated method. Due to the strict control and difficulty of the experim...

AI Technical Summary

Problems solved by technology

[0004] The cell membrane of HaCaT cells is relatively thick, difficult to digest, and even more difficult to transfect. It is very difficult to transfect foreign DNA into HaCaT cells through liposomes. The transfection efficiency is very low and often fails to meet the experimental requirements.

The efficiency of transfection by electric shock method and virus method is not high, and the experimental operation process is cumbersome. How to use simple liposome transfection method to improve the transfection of exogenous DNA into HaCaT cells has become a technical problem.

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