Monday, September 26, 2011

Enhanced Transfection Efficiency of Human Embryonic Stem Cells

Stem cell based drug discovery and cell based therapies hold great promise. Researchers are faced with many hurdles in developing and launching meaningful therapies. Key will be a thorough understanding of  the genetically stability of the stem cells involved in these therapies. Transfection tools will help foward this understanding. Here's a representative publication: Luis G. Villa-Diaz, Jose L. Garcia-Perez and Paul H. Krebsbach. Stem Cells and Development. December 2010, 19(12): 1949-1957. doi:10.1089/scd.2009.0505.

Overview: Because human embryonic stem (hES) cells can differentiate into virtually any cell type in the human body, these cells hold promise for regenerative medicine. The genetic manipulation of hES cells will enhance our understanding of genes involved in early development and will accelerate their potential use and application for regenerative medicine. The objective of this study was to increase the transfection efficiency of plasmid DNA into hES cells by modifying a standard reverse transfection (RT) protocol of lipofection. We hypothesized that immobilization of plasmid DNA in extracellular matrix would be a more efficient method for plasmid transfer due to the affinity of hES cells for substrates such as Matrigel and to the prolonged exposure of cells to plasmid DNA. Our results demonstrate that this modification doubled the transfection efficiency of hES cells and the generation of clonal cell lines containing a piece of foreign DNA stably inserted in their genomes compared to results obtained with standard forward transfection. In addition, treatment with dimethyl sulfoxide further increased the transfection efficiency of hES cells. In conclusion, modifications to the RT protocol of lipofection result in a significant and robust increase in the transfection efficiency of hES cells.

Related Neuromics' Reagents for Testing and Gene Expression Analysis of Stem Cells:
LumiSTEM™ in vitro Cell Based Assays-Designed for Primary Stem and Explanted Cells, Stem and Other Cell Lines.
LUMENESC™ in vitro Cell Based Assays-Designed for Mesenchymal Stem/Stromal Cells (MSCs) and Cells derived from MSCs.
HALO® in vitro Cell Based Assays-Designed for Lympho-Hematopoietic Stem and Progenitor Cells.
Transfection Reagents

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