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Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal,,,Fibroblasts


Improve transfection of primary cells up to 20-fold with Primary ENHANCER reagent

David Staggs
Genetic Applications LLC

Peter Pingerelli
Stratagene

Stratagenes Primary ENHANCER transfection reagent promotes efficient transfection of human dermal fibroblasts. Twenty minutes before transfection, cells are incubated in media containing the Primary ENHANCER reagent. After incubation, lipid or calcium phosphate (CaPO4)-mediated transfection can be performed using standard protocols. In earlier results, Primary ENHANCER reagent increased transfection efficiencies of some primary cell strains as high as 15-fold.1 In this study, we compared its effects on lipid- and CaPO4-mediated transfection using Stratagenes Normal Human Dermal Fibroblasts.

Human dermal fibroblasts are used to investigate many disease processes, including cancer, skin ulcers, collagen diseases, and scleroderma. They are also valuable for studying wound healing, growth factors, artificial skin, and dermal toxicity.

To study gene expression, a common approach usually involves introducing exogenous DNA into cells using transfection techniques in order to investigate the effects of resulting proteins. Unfortunately, primary human fibroblasts are difficult to transfect. However, with Stratagenes Human Dermal Fibroblasts, the method with which to study molecular and biochemical mechanisms in this system is improved. Additionally, Stratagenes new Primary ENHANCER reagent improves the transfection efficiency of lipid- or CaPO4-mediated transfection of human dermal fibroblasts.

Easily Increased Transfection

We grew Stratagenes Human Dermal Fibroblasts, which are isolated from adult skin and cryopreserved in vials (500,00 cells/vial),2 for two passages in standard
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