Fibroblast Transfection Information

Fibroblast cells from humans and other mammals are often used in biological research utilizing cell transfection methods. Some of the commonly used fibroblast cell lines are 3T3, a murine fibroblast line established in 1962 at the NYU School of Medicine. 3T3 cells were originally sourced from Swiss albino mice embryos and spontaneously immortalized. Another commonly used cell line is MRC-5, derived from human lung tissue.

Fibroblast Transfection

Transfection is the introduction of foreign molecules and genetic material into cultured cells. Transfections can be transient or stable, and the process is utilized in biological research to study gene function and to model the effect of gene therapy on cellular expression. In transient transfections, the short term impact of altered gene function is examined, with high expression persisting for 72 hours or less. Stable transfection requires that target genes be incorporated into the genome of a cell culture in such a way that expression continues throughout the life of the cell line and during each round of cell selection during subculturing.

pDNA and mRNA can be introduced via transfection, as well as siRNA and miRNA, which disrupt the development of gene products. This method of post-transcriptional gene silencing has a huge potential in terms of drug therapies that target genetic disease at the source, targeting the gene itself with a high degree of specificity, rather than relying on symptomatic or higher level treatment.

There are a great variety of carrier molecules on the market today which enable scientists to introduce genetic material into cancer or primary cells through non-viral means. This can be accomplished using immortalized cell culture lines, but also in vivo. However, all cells are different, and require different transfection reagents, carrier molecules, transfection protocols and reagents in order to successfully express a vector gene of interest. For links to resources for transfection and biological services, products and reagents, see below.

Links:

Fibroblast Transfection Kits – Purchase Fibroblast Transfection Reagent

Protocols, techniques, methods, in vivo transfection – siRNA Transfection

Xenograft.net – Preclinical biology CRO research services

Cell Transfection Resource

Transfection. Cells and Molecular Biology Research Methods, Protocols, and Lab Techniques

Biology Transfection Forum

Research Articles:

  • Similarities between fibroblasts and embryonic stem cells: This study compared MRC-5 fibroblast cells to embryonic stem cells in terms of morphology, growth rate and gene expression. In addition, the researchers developed a serum-free culture system that maintains the MRC-5 cells in a proliferative state similar to that of the embryonic cells. [LINK]
  • Response of fibroblast cell lines to hydrogen peroxide: Researchers studied the effect of hydrogen peroxide on human fibroblast cells, as peroxide has been previously shown to induce apoptosis or senescence in cultured cells. It was found that hydrogen peroxide in increasing concentrations produced a transient shock state, then senescence or apoptosis. [LINK]