Method for the Episomal Plasmide Vector
Here, we introduce the method of establishing iPS cells from a peripheral blood monocyte by introducing the initialization factor with the episomal plasmid. This is the low risk method for chromosome integration of exogenously introduced genes.. The feeder-free method is employed with the StemFit Basic medium and iMatrix-511.
(i) Preparation of the plate
(ii) Establishment by the episomal plasmid vector
Equipment/Reagent to be Used
Point of Operation
Take special care not to introduce air bubbles when transferring to the cuvette for electroporation.
[After gene transfer]
Immediately after introducing the gene to the cells by electroporation, suspend the cells in the medium previously warmed to 37°C in 5% CO2 incubator and inoculate onto a plate.
[After culture initiation]
To prevent flotation of the adherent monocyte, the StemFit® medium will be added carefully along the wall of the well.
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Establishment with Virus Vector