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View additional product information for Lipofectamine™ Stem Transfection Reagent - FAQs (STEM00001, STEM00003, STEM00015, STEM00008)
17 product FAQs found
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.
Yes. Lipofectamine Stem Transfection Reagent can deliver small and large (up to 11 kb) DNA plasmids into stem cells with very low cytotoxicity.
Certain components of culture media can be inhibitory to lipid-based transfections. For best results, we recommend using OptiMEM Reduced Serum Medium for preparing Lipofectamine Stem Transfection Reagent -DNA, -mRNA, and -Cas9 RNP transfection complexes.
Yes. Lipofectamine Stem Transfection Reagent is optimized for genome editing in human pluripotent stem cells and neural stem cells. Lipofectamine Stem Transfection Reagent provides efficient transfection of large DNA plasmids such as CRISPR/Cas9 vectors, RNA such as Cas9 mRNA co-transfected with tracr/guide RNA, or Ribonucleoprotein complexes of Cas9 protein with guide RNAs.
Proliferating stem cells in culture need room to expand. We recommended plating cells so they will be 30-60% confluent at the time 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.
When planning your transfection-based experiments, it is important to optimize the amount of nucleic acid and transfection reagent for each stem cell type being used, so that you may balance optimum transfection efficiency with minimal cytotoxicity.
We recommend using Lipofectamine RNAiMAX Transfection Reagent for siRNA delivery.
We recommend using our feeder-free Essential 8 Medium or StemFlex Medium on Vitronectin, Geltrex or Laminin-521 matrices. Lipofectamine Stem Transfection Reagent also supports other media systems such as mTeSR1 on Geltrex matrix.
Yes. Lipofectamine Stem Transfection Reagent has been tested with StemFlex Medium and Essential 8 Medium on Vitronectin, Geltrex, and Laminin-521 matrices.
Yes. High transfection efficiency can be achieved in stem cells in suspension with reverse transfection using Lipofectamine Stem Transfection Reagent. Stem cells can easily be transfected at the time of passaging and during re-plating, when they are in suspension. For simple instructions on using reverse transfection with Lipofectamine Stem Transfection Reagent, please take a look at the transfection protocols for pluripotent stem cells, listed under Documents, Product Literature on the product page (https://www.thermofisher.com/order/catalog/product/STEM00008?ICID=search-product).
Yes. Most albumin- and serum-supplemnted media do not inhibit transfection with this reagent.
Lipofectamine Stem Transfection Reagent can be used to deliver and co-deliver DNA, mRNA, and Cas9 protein (RNP) into stem cells, including neural stem cells (NSCs) and mesenchymal stem cells (MSCs).
Lipofectamine Stem Transfection Reagent is a lipid-based transfection reagent that is specifically designed for stem cells. It is our most versatile, and preferred choice for delivering DNA, mRNA, and Cas9 protein (RNP) into human pluripotent stem cells, as well as neural stem cells (NSCs) and mesenchymal stem cells (MSCs). Lipofectamine 3000 Transfection Reagent can be used to deliver DNA or RNA and is recommended for hard-to-transfect cell lines and workhorse cell lines.