Generation of Stable Cell Lines Using Retroviral Vector

Using retrovirus to generate stable cell lines has several distinct advantages over other vector systems:

1. Retroviral vectors have the ability to transform its single-stranded RNA (ssRNA) genome into a double-stranded DNA (dsDNA) molecule and stably integrate into a host genome at the transcriptional active site
2. All structural genes of the retrovirus are removed in the expression vector to prevent revertant recombination, making the retroviral vector safe and powerful tool for gene transfer
3. Retroviral vector-mediated gene transfer has been widely used in the development of gene therapy
4. Retroviral vectors are compatible with dividing cells and other primary cells
5. Retroviral vector based on moloney murine and leukemia viruse (Mo-MLV) is capable of infecting both mouse and human cells 
6. The viral genes (gag, pol, and env) in the retroviral system are replaced with the transgene and expressed in trans in the packaging cells

Fig. 1.  Generation of Stable Cell Lines by Retrovirus-mediated Gene Transfer Techniques

Stable Cell Pool Generation allows customers to obtain a stable cell pool for research in just four weeks (Fig.1). The ability to provide fast turnaround is based on the use of an artificial antigen that is expressed on the surface of transformed cells. Mammalian cells are transduced using modified retrovirus (Mo-MLV) vector(s) containing gene(s) of interest (Fig.2). The Mo-MLV transfer vector has all major structural proteins removed, rendering it incapable of replication or reversion. Once transduced, cells are allowed to grow for two weeks in a healthy pool. Then, transformed cells are sorted using specific antibody against the surface antigen (Fig.3). This technology provides a healthier cell pool since harsh antibiotics are never used. The established stable cell line is suitable for studying gene function, expression, toxicity, proliferation, as well as for small to medium scale production of recombinant proteins.

Stable Cell Line Generation service is designed for the manufacturing purposes, particularly for the production of recombinant antibodies in mammalian cells. An in-house engineered super activated expression vector system is employed. Highly expressed neomycin resistant clones are screened and isolated. The most highly expressed clones are selected, adapted to serum-free system, and developed as seed clones for downstream manufacturing.

Fig. 2.  Retrovirus Production in Packaging Cells

Fig. 3.  Cell Sorting To Select Integrated Cells

 

 Other advantages of our stable cell line include:

• Use of in-house developed retrovirus expression vector to ensure stable integration of your gene of interest
• Every project is designed and handled by scientists with over 20 years of experience
• Guaranteed >95% of cell population containing stably integrated gene for your protein
• Guaranteed the fastest turn around for widely used cell lines (Fig.4)
• Lowest price
• Detailed documentation of analysis

Fig. 4.  Timelines for Generating Stable Cell Lines

 Note: Some cells require additional testing that may increase the time required for production.

 

Back to cell line production services list