FIBROBLAST IMMORTALIZATION

Fibroblasts are morphologically heterogeneous and depending on their origin in the body display different functions. In addition to the widespread use in cosmetic research and skin models, human fibroblasts have emerged as a relevant target cell type for which involvement in complex tumor-stroma biology has recently been recognized.

To avoid batch to batch variations when working with primary fibroblasts and circumvent the restricted replicative capacity, immortalization is the method of choice.


Fields of Application

  • Oncology
  • Wound healing studies
  • Dermatological research
  • Tissue regeneration
  • Epithelial-mesenchymal interaction studies

Immortalization Strategy

SIRION BIOTECH has developed a lentiviral vector system for conditional gene regulation to immortalize primary cells with retention of terminal differentiation capabilities.

The thermosensitive variant of the large T antigen (large T (ts)) has already been successfully proven as immortalization gene for fibroblasts in a variety of experiments.

expression of this protein is comparable to a switch, having the large T antigen expressed as active protein at 33°C for cell proliferation and expansion. At 37°C largeT becomes inactive leading to a shift of cells closer to the primary status and thus enabling differentiation and functional studies.


Service details

  • Cultivation of primary cells
  • optional: characterization of gene expression profiles / long term cultivation
  • Transduction of primary cells with large T (ts) expressing lentiviruses
  • optional: any other immortalization gene/ gene combination
  • Generation of stable cell pools by antibiotic selection
  • Quantification of gene expression by qRT-PCR and QC
  • optional: on Western Blot
  • optional: characterization of gene expression profiles / long term cultivation
  • optional: clonal cell line generation
  • Delivery of frozen growing primary fibroblasts ready to use under BL-1

NOTE: SIRION Biotech cannot guarantee any retention of functional and morphological primary cell characteristics using this standard immortalization approach. Contact us for more information.


Primary cell origin

Primary cells are supplied by the customer and tested

  • to be free of human pathogen viruses (HCV, BCV, and HBV)
  • for the absence of contamination with mycoplasma.