Immortalization of eukaryotic cells
In researchers’ work, there is often a need to obtain permanent cell lines from primary cells. For example, when studying various human diseases, a sample of venous blood or a tissue biopsy can be taken from a patient, and a short-lived cell line can be generated from this material. Such cells are typically used for a wide range of studies. In particular, they can be used to develop personalized medicine approaches for a given patient, to produce large amounts of biomaterial for DNA extraction and for series of whole-genome sequencing experiments, and so on. For all of these applications, obtaining an immortalized line offers a number of advantages, as it removes limitations on the amount of material. In addition, a permanent cell line - serving as a model of a particular disease - can be derived from patients’ primary cells.
Various approaches are used to immortalize cells, and the choice depends on the type of cells to be immortalized as well as on the aims and objectives of the experiment (Fig. 1A).
Various approaches are used to immortalize cells, and the choice depends on the type of cells to be immortalized as well as on the aims and objectives of the experiment (Fig. 1A).
Figure 1. (A) Methods for immortalization of eukaryotic cells. (B) Generation of immortalized chicken embryonic fibroblasts. [doi.org/10.1038/s43016-022-00658-w].
The simplest approach is to obtain spontaneously immortalized cells. For example, this approach was used to develop a method for producing cultivated chicken meat from chicken embryonic fibroblasts [doi.org/10.1038/s43016-022-00658-w]. In that work, during prolonged cultivation, colonies of immortalized cells were obtained within an aged, dying culture (Fig. 1B). Immortalization of the resulting cells was confirmed based on the expression profile of relevant genes, and then, by scaling up biomass production, cultivated meat was produced. The spontaneous immortalization method makes it possible to obtain passaged cell lines without any genetic manipulation by the researcher; however, it requires a very long time and provides no guarantee that immortalization will occur.
One of the most efficient immortalization approaches is based on the use of conditioned culture medium in which cells infected with Epstein–Barr virus, one of the most common human viruses, were cultured [doi.org/10.1007/BF02633976]. A major advantage of this method is that in some cases the immortalization efficiency reaches 98% and no large chromosomal rearrangements occur. However, a drawback of the technique is that it is applicable only to certain cell types that are susceptible to the virus.
Another method with very high efficiency and broad applicability is based on delivering into cells a transgene encoding either the catalytic subunit of telomerase or the SV40 large T antigen. Using both transgenes simultaneously can also be highly effective [doi.org/10.1038/sj.onc.1205014]. The choice of transgene depends on the type of cells to be immortalized and on the species from which the cells are derived. The limitations of this approach are primarily related to the cells’ susceptibility to foreign DNA - that is, how efficiently they can be transfected. At the same time, it remains possible to immortalize poorly transfectable cells due to the high immortalization efficiency of the transgene.
Depending on the type of cells that need to be immortalized, the method most effective in a given case is selected.
Our company offers immortalization of:
-Human peripheral blood B lymphocytes.
-Fibroblasts from any mammals and birds.
-Other cell types, for which we will select the most effective method.
To carry out immortalization, you need to contact our specialists, and in consultation with you we will select the most optimal method that will allow us to obtain a passaged cell line for you in the shortest possible time. Upon completion, you will receive live cells in the quantity you require and, if needed, information on gene expression profiles and immortalization markers.
One of the most efficient immortalization approaches is based on the use of conditioned culture medium in which cells infected with Epstein–Barr virus, one of the most common human viruses, were cultured [doi.org/10.1007/BF02633976]. A major advantage of this method is that in some cases the immortalization efficiency reaches 98% and no large chromosomal rearrangements occur. However, a drawback of the technique is that it is applicable only to certain cell types that are susceptible to the virus.
Another method with very high efficiency and broad applicability is based on delivering into cells a transgene encoding either the catalytic subunit of telomerase or the SV40 large T antigen. Using both transgenes simultaneously can also be highly effective [doi.org/10.1038/sj.onc.1205014]. The choice of transgene depends on the type of cells to be immortalized and on the species from which the cells are derived. The limitations of this approach are primarily related to the cells’ susceptibility to foreign DNA - that is, how efficiently they can be transfected. At the same time, it remains possible to immortalize poorly transfectable cells due to the high immortalization efficiency of the transgene.
Depending on the type of cells that need to be immortalized, the method most effective in a given case is selected.
Our company offers immortalization of:
-Human peripheral blood B lymphocytes.
-Fibroblasts from any mammals and birds.
-Other cell types, for which we will select the most effective method.
To carry out immortalization, you need to contact our specialists, and in consultation with you we will select the most optimal method that will allow us to obtain a passaged cell line for you in the shortest possible time. Upon completion, you will receive live cells in the quantity you require and, if needed, information on gene expression profiles and immortalization markers.