Flash Therapeutics

GENERATION OF STABLE CELL LINE USING LENTIVIRAL VECTORS

Vectalys provides a custom stable cell line generation service for many applications. The use of lentiviral vectors for stable cell line generation allows for optimal transduction of immortalized, primary and stem cells.

In delicate cells such as hematopoietic lineages for example, classical transfection protocols are not only transient and inefficient, they also induce cytotoxicity effects and proliferative arrests. Gene transfer using highly concentrated and purified lentiviral vectors allows for efficient and safe transduction of such cells.

Lentiviral vectors : a tool of choice for cell engineering

Lentiviral vectors are the engineering tool of choice to study and engineer primary cells without interfering with the cell phenotype, and are also the only tool capable of transducing primary cells without toxicity.

Additionally, the advantages of lentiviral vectors combined with our new technology LentiFlash allow for a faster expression without genomic traces in the target primary cells.

This make Vectalys’ lentiviral vectors the ideal tools for gene editing, cell differentiation, and immunotherapy.

CUSTOM STABLE CELL LINES

Vectalys generates customized stable cell lines from immortalized to primary and stem cells.
Since the creation of the company in 2005, Vectalys has ran more than 1200 cell line generation projects using highly pure and concentrated lentiviral vectors. When considering gene transfer into mammalian cells, the purification level of lentiviral vectors is critical. Vectalys has developed several specific purification levels according to the target cells.

Maintain the original cell phenotype

Using highly pure lentiviral vectors, Vectalys provides stable cell lines while maintaining the original phenotype to mimic, as much as possible, natural in vivo conditions. For example, transduced T cells exhibit an original cell phenotype without any changes to T cells specific marker expression such as CD44, CD69, CD25 and Ly-6C, indicating that the use of our highly pure lentiviral vectors does not impact the T cells’ phenotype.

Manage and control expression level

Cell lines are commonly used for assay development and screening. One of the issues using such cell lines is controlling transgene expression. Being able to fully characterize expression rate and the number of copies in cells is essential to design robust screening assays.

In classical transfection approaches, the challenge is to ensure an appropriate and controlled expression level in target cells for any transgene. Thanks to high quality lentiviral vectors, Vectalys offers the opportunity to observe a quantitative effect linked to a differential gene expression level, by using a controlled Multiplicity of Infection (MOI).

Eliminate the need for antibiotic resistance

Antibiotics can influence the physiology of your target cells which is harmful when working with primary cells to maintain accurate natural in vivo conditions. Because it is possible to achieve 100% of transduced cells in only one experiment by using lentiviral vectors, the use of selection markers such as antibiotics is not required.

Save time and cost

Most of the transfection issues are about gene transfer efficiency into hard to transfect cells (lymphoid and myeloid cell lines, primary and stem cells). Using lentiviral vectors, the expression of the sequence of interest (cDNA, shRNA, miRNA) is stable thanks to the vectors DNA integration into the host genome, providing a permanent cell line in one experiment. This avoids having to transfect targeted cells multiple times for bioproduction or screening (CHO, HEK…). Lentiviral vectors allow to save time, money and energy.

Custom stable cell lines expressing your gene of interest

Obtain your custom stable cell lines with lentiviral vectors dedicated to your target cells. Discover our many possibilities on any cell type:

Immortalized cell lines

• Human cell lines
• Mouse cell lines
• CHO cells

Primary cells

• Monocytes
• T/B Lymphocytes
• Dentritic cells
• Neurons/Astrocytes
• Oligodendrocytes
• Endothelial cells
• Tumoral cells
• Hepatocytes
• Muscle cells

Stem cells

• Hematopoitetic SC
• Mesenchymal SC
• iPS cells
• ES cells

Lentiviral vectors are time saving tools for stable cell line generation. Using Vectalys Lentiviral vector tools you will be able to get from your gene of interest to the polyclonal cell line generation in a month.

The viral vectors provided by Vectalys, by virtue of their high titre and purity, minimize the deleterious phenotypic changes that typically occur following transduction of target cells, such as loss of a sub-populations of transduced cells, and effects on proliferation, viability and differentiation of transduced cells.

Benefits of Vectalys lentiviral vectors
Vectors produced by Vectalys present 3 main advantages based on the absence of serum and on a reliable titer that:

• Ensure that the integration events occurs in every cells at a specific MOI– 100% transduction efficiency
• Enable management and control of the M.O.I. (dose effect)
• Eliminate needs for antibiotic resistance selection steps

Vectalys specific serum free purified vectors batches (C) for primary cells transduction show several advantages:

• The Vectalys patented lentiviral vectors do not affect the viability and the proliferation of your target cells
• Vectalys lentiviral vectors enable to manage both % of positive cells and gene of interest (G.O.I.) level expression by controlling the M.O.I.
• Vectalys lentiviral vectors ensure that all the target cells are genetically modified – 100% transduction efficiency on primary cells and stem cells.
• Vectalys lentiviral vectors eliminate needs for antibiotic resistance selection steps

Vectalys lentiviral vectors for in vivo models

In vivo, lentiviral vectors are efficient tools for noninvasive analysis of tumor animal models. They can be used through direct injection into adult animals or through in vitro transductions of any tumoral cells subsequently reimplanted in the context of cancer models.

Lentiviral vectors have to be highly concentrated and purified in order to achieve an effective transgene expression while maintaining the cells original phenotype, proliferation and viability. These criteria for the development of trustworthy and predictive cancer models are fulfilled by the properties of our lentiviral vectors, building a bridge between in vitro assays, in vivo results and finally therapeutic applications.