The day before transfection, trypsinize and count the cells plate 4 x 10 4 cells per well in 05 ml of complete growth medium cell density should be 50~80% confluent on the day of transfection for each well of cells to be transfected, dilute 05 μg of dna into 100 μl of opti-mem® i reduced serum medium without serum.
Background primary brain capillary endothelial cells (bcecs) are a promising tool to study the blood–brain barrier (bbb) in vitro, as they maintain many important characteristics of the bbb in vivo, especially when co-cultured with pericytes and/or astrocytes. I use sheep endothelial cells and have struggled for years but now i can transfect them without too much trouble you need to give it a try to figure out only one concern is the fact that mouse ec primary culture usually can not be be expanded so i do not know whether you can have enough ec for your transfection.
Transfection of endothelial cells essay sample transfection for hmecs using lipofectamine ltx + plus reagent nb: • using plus™ reagent (cat no 11514-015) enhances transfection performance in huvec cells. For efficient transfection of specific primary cells, eg mouse b-cells or mouse immature dcs, in the 4d-nucleofector tm x unit (100 µl format.
Primary cells are considered more difficult to transfect than immortalized cell lines, as they are more susceptible to toxic agents and may degrade exogenous nucleic acids in the cytoplasm 1, 2 in vitro genetic modification of primary endothelial cells, such as huvec, is used in the study of gene function, 3 angiogenesis, 4 and for applications in gene therapy, 5 among others. Transfection, the introduction of nucleic acid into cells, is a powerful technique used to study in vivo gene function and regulation the cytofect™ endothelial cell transfection kit is a plasmid dna delivery system specifically optimized to deliver dna into a wide variety of endothelial cells.
Transfection of brain capillary endothelial cells in primary culture with defined blood–brain barrier properties annette burkhart 1 , louiza bohn thomsen 1 . Cell density should be 50~80% confluent on the day of transfection (use the normal growth medium without antibiotics) 2 for each well of cells to be transfected, dilute 1 μg of dna into 200 μl of opti-mem® medium without serum.
For efficient transfection of specific primary cells, eg mouse b-cells or mouse immature dcs, in the 4d-nucleofector tm x unit (20 µl format.