Cell Line Services

Custom Stable Cell Pools

In cooperation with SIRION Biotech, ibidi offers the following custom stable cell pools that can be used in various applications:

LifeAct–GFP / RFP Expressing Cell Pools:

These cell pools allow in vivo imaging of the actin cytoskeleton and permit the study of actin dynamics by using the advantageous LifeAct actin marker.

Bright field and fluorescence images of stably transfected Normal Human Dermal Fibroblasts (NHDF) expressing LifeAct-RFP.

Cell Pools for Protein Overexpression:

These cell pools allow the stable expression of your gene of interest in your mammalian cell line of interest for use in:

• Purification of proteins
• Functional gene analysis
• Target identification and validation
• Generation of reporter cell lines, e.g., for screening
• Expression of fluorescent or luminescent proteins for in vivo imaging

Western Blot showing the expression of two splice variants of a glycolytic enzyme in stable THP-1 cell pools (Lanes 4 and 5), both of which are not expressed in the parental cell line (lanes 3). Lanes 1 and 2 show positive controls.

Cell Pools for Simultaneous Expression of Two
or More Proteins:

These cell pools allow stable mammalian expression of your genes of interest in your cell line of interest for use in:

• Analysis of protein/protein interaction
• Expression of heteromeric proteins
• Generation of reporter cell lines, e.g., for screening
• Coexpression of fluorescent or luminescent proteins for FACS analysis or in vivo imaging (xenotransplant)

Services available with custom stable cell pools include:

• Cloning of the cDNA from your gene(s) of interest into an expression vector*
• Verification of cloning success by DNA-sequencing*
• Transfection of your cell line
• Generation of stable cell pools by antibiotic selection
• Expression quantification of your gene of interest using qRT-PCR and/or Western Blot, or fluorescence
• Delivery of validated frozen cells

* service not needed when using LifeAct GFP/RFP

FACS of Stable Cell Pools

If you want to improve the homogeneity of your cell pool using fluorescent or luminescent proteins, ibidi provides a cell sorting service based on flow cytometry.

This additional service comprises:

• FACS of a stable cell pool to isolate specific subpopulations
• Customized expansion and cell banking of sorted cell pools

Stable murine 4T1 cell pool simultaneously expressing RFP and GFP from two different vectors.

Histograms showing a stable HT-1080 cell line, generated using LifeAct-TagGFP2, before (middle) and after (top) FACS in comparison to HT-1080 wt cells (bottom). Through the use of FACS, a homogeneous cell population with more than 95 % of fluorescent cells was obtained.

Clonal Cell Line Service

For customers who need optimal transgene expression and guaranteed stability of cell clones, ibidi provides a Clonal Cell Line Service.

This service includes:

• Establishment of single cell clones using the Genetix CloneSelect Imager
• Small scale expansion of individual clones
• Identification of „High Performers“ via expression quantification of a target gene, or gene of interest, using qRT-PCR and/or Western Blot, relative to control cells
• Large scale expansion and cell banking of selected clones
• Final quality control of transgene expression, growth characteristics, and long-term cultivation (20 passages)
• Optional: marker gene expression profiling, genotyping, and genomic mapping of transgene integration site

Caco-2 cells are used as an in vitro model of the human small intestinal mucosa to predict the absorption of orally administered drugs. In order to understand mechanisms
of drug efflux better, SIRION BIOTECH generated a Caco-2 cell line with stable knockdown of P-Glycoprotein 1 (pgp1) of more than 80 % over 20 passages.

Functional Cell Characterization

If you want to enhance the reliability and authenticity of your stable cell line (pool or clone) ibidi provides a functional characterization service that uses ibidi’s versatile cell-based assays.

The Functional Characterization service includes the following assays (when appropriate):

• 2D and 3D Chemotaxis
• Wound Healing
• Angiogenesis
• Cell Culture under Flow
• Genomic Analyses (carried out by the DSMZ)
• Cell Proliferation Analysis

The HT-1080 cell line is a tumor-model system widely used in vitro and in vivo. Here, a stable HT-1080 cell line was analyzed using a chemotaxis 3D assay, with wt HT-1080 cells being used as a control.The analysis revealed that there was no significant difference in chemotaxis and velocity in the two cell lines.