What is the difference between transient and stable cell lines?

Transient cell lines are those in which the target gene is not permanently integrated into the host cell genome, resulting in gene expression occurring only for a limited period. Stable cell lines contain target genes that are integrated into the host genome to support the long-term expression of target genes.

Is the time and expense necessary to build a stable cell line?

If trial and error is required on cells that overexpress or reduce the expression of the desired gene, then the construction of stable cell lines is necessary. First, it can reduce the difference in experimental results caused by the difference in transfection efficiency in different batches of experiments, and improve the repeatability of the experiment. Second, after the construction of the cell line, there is no need to perform instantaneous transfection experiments and repeated preparation of plasmids, reducing the workload. Thirdly, expensive transfection reagents are no longer used to save experimental costs.

What are the advantages of stable cell lines?

Key advantages of stable cell lines include significantly reduced variability between production batches, applicability to a wide range of research and production applications, and robust growth performance.

How many generations can the target gene be stably expressed?

The characteristics of the lentiviral system determine that the target genes are always expressed like the cell’s own genes, and are not lost in the process of passage.

What is the difference between monoclonal and polyclonal cell lines?

Monoclonal cells are cloned and screened after transfection of the target gene. The cell line formed by the expansion of one cell has the same integrated location and copy number of the target gene, and the expression level can be maintained for a long time. Polyclonal cells were directly screened by a drug resistance gene after transfection of target gene. It’s a mixture of multiple positive cell clones. For information on monoclonal and polyclonal cell lines, see our monoclonal cell Lines Introduction.

How long does it take to produce a stable cell line?

Developing a stable cell line typically takes 6 to 12 months, and this time frame varies depending on the complexity of the specific target protein or biologics being pursued.

Can stable cell lines be constructed from all cell lines?

Not all cell lines can be constructed into stable cell lines. Two conditions need to be met to construct stable cell lines: 1. Stable cell passage; 2. The transfection efficiency of cells met the requirements. We offer a free pre-lab service to test whether your cell line can be used for stable cell line construction.

How to choose the right lentiviral vector?

The selection of lentiviral vectors mainly requires consideration of promoter selection, whether fluorescent protein tags are required, and whether fluorescent protein tags are required for fusion expression. Commonly used lentiviruses are pLV-puro, pLV-EGFP-C, pLV-sfGFP (2A) puro.

What is the method for constructing stable cell lines?

Stable cell lines were constructed by lentivirus transfection. The characteristic of this system is that the target gene is first integrated into the genome and then expressed, without instantaneous expression process. Therefore, the screened cells are stable and integrated.

References

[1] Tihanyi B, Nyitray L. Recent advances in CHO cell line development for recombinant protein production. Drug Discov Today Technol. 2020 Dec;38:25-34.

[2] Chen LQ, Wang YN, Li DY, Xu LY, Li LJ, Li ZH, Liu H, Liu Q. Construction of a Stable Expression Cell Line of Human Phospholamban. Fa Yi Xue Za Zhi. 2021 Oct 25;37(5):615-620.

[3] Lü X, Zhou Z, Zhu L, Zhou J, Huang H, Zhang C, Liu X. [Construction and identification of a HEK293 cell line with stable TrxR1 overexpression]. Nan Fang Yi Ke Da Xue Xue Bao. 2022 Apr 20;42(4):554-560.