Primary mouse embryonic fibroblasts (MEFs) have a number of properties that make them an attractive cell culture model. Relative to other primary explant cultures they are easy to establish and maintain, proliferate rapidly and, as a result, large numbers of cells can be produced from a single embryo within several days following explanation. This allows, for instance, for ready comparison of wild-type and knockout cells derived from the same litter of animals. MEFs can be expanded through several passages before they reach crisis and can be used to establish cell lines following spontaneous transformation or following derivation from strains carrying mutations, such as in the gene encoding the tumor suppressor Trp53. They have been widely used as feeders to support other cultured cell types, notably embryonic stem cells, as well as for the study of a diverse range of cellular phenomena using microscopic, biochemical and molecular biological techniques.
The Transfection Reagent:
Transfection of primary cells, such as MEFs, has always been challenging, both technically and financially. Most methods working effectively on cell lines cause significant toxicity on primary cells, thus fail to transfect efficiently.
MEF Avalanche® Transfection Reagent (Mouse Embryonic Fibroblast) is a new class of unique chemical formulations specifically formulated and optimized for transfecting MEFs. The proprietary formulation of lipids and polymers ensures the highest possible transfection efficiencies and viabilities for MEFs. No other transfection reagents can match the efficiency, convenience, and gentleness of MEF Avalanche® Transfection Reagent for transfection on MEFs. For details of the developing process of this reagent, please go to: How did EZ Biosystems develop Avalanche® Cell type/cell line specific transfection reagent series?
- Specifically optimized to deliver nucleic acids into MEF
- Highest efficiency to ensure experimental success
- Extremely gentle to cells
- Deliver single or multiple plasmids
- synthesized from 100% animal origin-free components, making it easy to validate the absence of zoonotic diseases, such as BSE or viruses, in research experiments or cells lines
- Simplest and straightforward protocols
- Compatible with serum
- Reproducible: due to highly controlled chemical synthesis of each of the ingredients, the reagent forms uniformly sized complex particles with nucleic acids. With optimized protocol, our reagent will ensure the reproducible highest transfection results.
- Economical: High efficiency means less amount of nucleic acid & reagent is needed
- Developed and manufactured by EZ Biosystems
|FIG. 1. High throughput test of transfection efficiency (determined as RLU/mg) on MEFs after transfection of luciferase reporter gene by using our 172 proprietary transfection formulas and several most popular commercial transfection reagents. The yellow box showed the results of 4 commercial transfection reagents. The red lines marked our candidate formulas with the highest transfection efficiency for MEFs. This test result was confirmed with repeat experiments. The one that showed the optimal balance of potent & low cytotoxicity among those candidate formulas after flow cytometry analysis on the percentage of 7AAD positive cells was later named as MEF Avalanche® Transfection Reagent.
|FIG. 2. MEF cells were transfected with GFP vector (pEGFP-N3) by using MEF Avalanche® Transfection Reagent. The cells were visualized by Nikon Eclipse Fluorescence microscope 24 hours post transfection.
For Other Cells:
|MEF Avalanche® Transfection Reagent (Mouse Embryonic Fibroblast) can also be used on the following cells with high transfection efficiencies.
Recommended protocols for these cells will be provided with the reagent. The protocols usually provide satisfactory transfection efficiency with invisible cytotoxicity. However, optimization may be needed for certain type of cells. Optimizations may include: the amount of DNA and this transfection reagent; cell density; transfection reagent/DNA ratio, or incubation time for the mixture of transfection reagent/DNA etc. For best transfection result, we recommend using the respective cell type/cell line specific Avalanche transfection reagents. Those reagents have been optimized on both recipes and protocols, and have been proved to have the best transfection results for the respective cell lines or primary cells. You can easily find the respective Avalanche transfection reagents specific for your cells by using the filters of our product list.
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