Xfect Mouse Embryonic Stem Cell Transfection Reagent
- Transfect mouse embryonic stem cells (mESC)
- Unmatched efficiency: up to >90%
- Achieve high levels of gene expression
- No cytotoxicity, maintains pluripotency status
- Simple, serum-compatible protocol
Xfect mESC Provides High Transfection Efficiency in mES Cells
Effective transfection of embryonic stem cells with Xfect mESC Transfection Reagent. ES-E14TG2a mES cells were transfected with DsRed-Express2 plasmid using Xfect mESC Transfection Reagent. 48 hr posttransfection, transfection efficiency was assessed via flow cytometry (A) as well as phase (B) and fluorescent microscopy (C). As quantified by flow cytometric analysis, 93.9% of the cells transfected with Xfect mESC Transfection Reagent were positive for DsRed-Express2 expression (A), with a mean fluorescence intensity of 716.9 (not shown).
Xfect mESC reagent offers higher transfection efficiencies
Xfect mESC Transfection Reagent provides superior transfection efficiency in mES cells. Panel A. Xfect mESC Transfection Reagent and three competitor reagents were used according to their respective protocols to transfect the cell lines listed above using plasmid DNA encoding AcGFP1, in a 6-well format. 48 hr posttransfection, AcGFP1 expression was assessed by flow cytometry in order to
determine transfection efficiency. Panel B. Xfect mESC Transfection Reagent and two competitor reagents were used according to their respective protocols to transfect ES-E14TG2a mES cells using plasmid DNA encoding secreted alkaline phosphatase (pCMVSEAP), in a 6-well format. 48 hr posttransfection, the amount of SEAP expressed was measured in order to calculate the relative
transfection efficiency (Panel B).
Xfect mESC Transfection Reagent Does Not Affect mES Cell Pluripotency
Xfect mESC Transfection Reagent does not alter the pluripotency status of mouse embryonic stem cells. D3 mES cells were transfected with reaction buffer alone, Xfect mESC Transfection Reagent polymer and reaction buffer, or Xfect mESC Transfection Reagent polymer and reaction buffer plus plasmid DNA encoding DsRed-Express2. 48 hr posttransfection, total RNA from each sample was collected and expression levels of five genes was determined by QRT-PCR: Nanog and Oct4 (pluripotency markers), Gata6 (endodermal marker), Brachyury (mesodermal marker), and Map2 (ectodermal marker). The data show that Xfect
mESC Transfection reagent did not induce nonspecific differentiation or alter the pluripotency status of D3 mES cells, as the expression level of these markers remained unchanged in transfected samples.
Simple Transfection Protocol that is Serum-Compatible
Xfect mESC Transfection Reagent forms biodegradable nanoparticles which provide a new and highly efficient means of introducing exogenous DNA into mammalian cells, using an easy transfection protocol. The Xfect mESC Transfection Reagent protocol requires few steps and minimal optimization, and is easily scalable. Transfections can be carried out entirely in the presence of serum.