Lipofectamine™ 干细胞转染试剂

To obtain the highest percent of stem cells transfected with Lipofectamine Stem Transfection Reagent, we highly recommend that you follow cell culture conditions that promote formation of a monolayer versus clumps of cells. If the density of your stem cells is greater than 50% or they are being grown as colonies in other media, more Lipofectamine Stem Transfection Reagent may improve transfection efficiency. For additional details, please refer to the Lipofectamine Stem Transfection Reagent protocol pertinent to your cell type and media on the product page (https://www.thermofisher.com/order/catalog/product/STEM00008?ICID=search-product), under Documents, Product Literature.

Yes. Lipofectamine Stem Transfection Reagent can be used to co-deliver multiple payloads, including DNA, mRNA, and Cas9 protein (RNP) into stem cells.

Cell confluency can impact the amount of Lipofectamine Stem Transfection Reagent that is needed for efficient transfection. Stem cells that are near confluency do not transfect well. Plating density should be such that cells are not overgrown 24 hours after transfection. To obtain the highest percent of stem cells transfected, we highly recommend that you dissociate both colony type and monolayer cultures into small clumps of 3-5 cells. If you are growing human pluripotent stem cells in a 24-well plate in Essential 8 Medium on Vitronectin, plate the cells to achieve 30-60% confluency on the day of transfection. For additional details, please refer to the Lipofectamine Stem Transfection Reagent protocol pertinent to your cell type and media on the product page (https://www.thermofisher.com/order/catalog/product/STEM00008?ICID=search-product), under Documents, Product Literature.

Yes. Lipofectamine Stem Transfection Reagent can be used to transfect a wide range of stem cell types, while supporting continued proliferation without inducing differentiation. It is our recommended solution for transfecting neural stem cells.

For cells that have aggregated into large colonies, transfection attempts may result in ONLY the outer cells of the colonies being successfully transfected. Transfection in suspension at the time of passaging and re-plating can maximize access of the reagent to all the cells and increase transfection efficiency. We recommend cell culture conditions that promote formation of a monolayer versus clumps of cells. To avoid cell aggregates that form large colonies, gently dissociate colonies to small clumps of 3-5 cells to promote efficient transfection. Follow recommended seeding densities for the cell culture medium and passaging reagents used so that cells are at the recommended starting confluency of 30-60% at the time of transfection and have room to grow for 24-48 hours during transfection.