Monoclonal cell culture method

Abstract

The application of the invention belongs to the technical field of culture of monoclonal cells, and particularly relates to the application matters of the invention of a monoclonal cell culture methodin the process of constructing a stable expression cell system through the gene editing technology. The method takes the gene editing technology CRISPR / Cas9 as the basis. The method specifically comprises the following steps: editing a genome through the CRISPR / Cas9 technology, and culturing a screened cell through a 3D soft fibrous protein matrigel. In a word, the culture condition of the optimized 3D soft fibrous protein matrigel is used to construct a monoclonal cell population, the survival rate of the single cell is greatly increased, and the stable transfection cell system sharing identical gene modification and identical genetic background is rapidly screened. Particularly for the cell difficult to growth through the common culture method, the monoclonal cell culture method has obvious advantages and is favorable for the implementation of the follow-up experiments.

Description

technical field [0001] This application belongs to the technical field of monoclonal cell culture, and specifically relates to a patent application for a monoclonal cell culture method in the process of constructing a stable transfected cell line using gene editing technology. Background technique [0002] The cell group formed by the multiplication of a single cell is called a monoclonal cell group, that is, the process of culturing a cell to make it divide continuously to form a cell group, which is derived from a common ancestor cell. With the use of lentiviral transfection technology to construct stable expression cell lines and the popularization of gene editing technology, the method of constructing monoclonal cell populations has also been developed. Especially in tumor-related research, how to obtain single and stable cell lines is particularly important. important. [0003] Lentiviruses belong to the retroviridae family and are RNA viruses, the best known of which ...

AI Technical Summary

Problems solved by technology

However, in the actual operation process, limited by factors such as cell type or cell viability, some single cells hardly grow, the cells have not divided, or the surviving single cells are extremely weak when the partially diluted cells are cultured on the 96-well plate. There are few problems, such as the need to spread multiple 96-well plates for culture to carry out subsequent screening operations. These defects either seriously affect subsequent experimental operations, or lead to unstable screening results. Therefore, it is extremely necessary to further improve the single-cell culture system

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