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

growth media. The cells were then plated in 24-well format tissue culture dishes and allowed to attach for 18 hours. The media was replaced with media containing 0, 10, or 40 M of Stratagenes Primary ENHANCER reagent. After 20 minutes, the cells were transfected with a plasmid expressing the b-galactosidase reporter molecule under the control of the cytomegalovirus promoter (pCMVbGAL). Transfection was performed using either Stratagenes LipoTAXI transfection reagent3 or a CaPO4-mediated transfection technique.4

The Primary ENHANCER reagents effects on lipid-mediated transfection show that expression of the reporter molecule improved over 5-fold after incubating in 10 M Primary ENHANCER reagent and 20-fold after incubating in 40 M Primary ENHANCER reagent (Figure 1). Results were measured by Stratagenes fluorescent b-galactosidase assay kit.

Figure 1

The Primary ENHANCER reagents effects from using a CaPO4-mediated transfection technique show that expression of the reporter molecule, b-galactosidase, improved slightly after incubating in 10 M Primary ENHANCER reagent and nearly 3-fold after incubating in 40 M Primary ENHANCER reagent (Figure 2). Results were measured by Stratagenes fluorescent b-galactosidase assay kit.

Figure 2

Conclusions

Use Stratagenes Human Dermal Fibroblasts to study molecular and biochemical mechanisms in human fibroblasts. We demonstrated that the transient transfection of fibroblasts improved up to 20-fold in lipid-mediated transfection and
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