A convenient and improved selection method for immortalisation of neural cells

Abstract

Abstract There is a wide variety of cells in the nervous system that collaborate with each other for information transfer, regulation and neuroprotection. Various cell investigations involving primary cells isolated from mammalian tissues are helpful in understanding disease pathophysiology. However, these cells are difficult to acquire in vitro in a short time and die of aging after repeated passages. These hurdles have hindered cell and disease research. Immortalized cell lines are necessary for cell research. After creating immortalized cell models, antibiotics are commonly employed to select immortalized cells. However, immortalized genes have poor transfection rates and a considerable percentage of non-immortalized cells are destroyed, shed, or are not harvested from suspension following antibiotic selection. Cells form a network of a variety of cell junctions. Consequently, when a large number of cells die and fall off, immortalized cells that were successfully transfected and had antibiotic resistance can also be removed, leaving only a small number of viable immortalized cells. Because of reciprocal nourishment between cells, when a few survive, their proliferation ability is also limited due to a lack of other cell secretion factors. Thus, obtaining immortalized cell lines takes more time. In this study, cells with transfected immortalized genes were cultured, subcultured, and selected. This process could generate stable immortalized cell lines in less time by enhancing the classic immortalized cell selection procedure. The feasibility and advantages of this method were demonstrated by infecting Schwann cells with retroviruses containing plasmid SSR#69 packaged in HEK293 cells. The resulting immortalized Schwann cell lines were compared to cells obtained by the standard screening before culture.