PerkinElmer and Oncosec will have a Patient-Selection Biomarker in Immunotherapy.
Cell-based therapy is currently at the forefront of tissue engineering strategies and is expected to dominate the field in the future. A recent study from the University of Hong Kong was aimed to optimize electroporation parameters for the transfection of human dental pulp stem cells, with cellular viability taken into account. Other chemical transfection agents were used for comparison, and their transfection efficiency and relevant cell viability were assessed.
The study compared the transfection of dental pulp stem cells (DPSCs) to a positive control using the commonly utilized chemical transfection reagents Lipofectamine 2000 and FuGene 6. From the results, they were able to obtain a higher transfection efficiency and cell viability with electroporation conditions compared to controls. The highest transfection efficiency (63.81±4.72%) was achieved with 100V, 20msec, one-pulse square-wave condition. Among the chemical transfection groups, FuGene 6 showed the highest cell viability at all tested transfection ratios, while Lipofectamine 2000 showed the highest transfection efficiency (19.23±3.19%) using 1:1 DNA
An increase in various chondrogenic markers was also found when studying mRNA expression in transfected cells. In conclusion, the results of the study demonstrated optimal electroporation and chemical transfection reagent conditions for human DPSCs, and, subsequently, provided proof of concept for expression of a functional gene using those conditions.
Now what does it have to with PerkinElmer you might ask. FuGene 6 Transfection Reagent has been commercially available since 1997 and since that time, its popularity has increased due to its ease of use, minimal to no cytotoxicity, and the high level of transfection in many different cell lines. It is used by PerkinElmer with their Alpha assays involving tagged recombinant proteins, which are transfected into cells. The approach is a preferred one due to the high protein expression levels achieved in transfected cells, and the availability of good anti-tag antibodies.
Many different technologies are available to transfect the expression vectors into cells (Calcium Phosphate, Electroporation, Ballistic Particles, DEAE Dextran, Cationic Matrix, and Lipofection). The lipofection protocol is a relatively simple method that has been used for high throughput screening. FuGENE6 has been observed to work best in the presence of serum and resulting in little or no toxicity. Only until recently results have proven that Electroporation has an advantage over other technologies, and PerkinElmer would probably prefer to use it with their assays.
“The analytical needs of both UCLA and OncoSec are well aligned with our unique technical capabilities,” said Brian Kim, President, Life Sciences & Technology, PerkinElmer. “This project complements our focus on enabling research to reveal the intricacies of immuno-oncologic interactions. Through this work with UCLA and OncoSec, we think we can make a meaningful contribution toward assisting researchers in improving outcomes for patients.”
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