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Invitrogen™ Lipofectamine™ Stem Transfection Reagent
Description
Lipofectamine Stem Transfection Reagent is optimized to achieve maximum efficiency with minimal early differentiation in a wide range of stem cells. It can co-deliver DNA, RNA, and Cas9 ribonucleoprotein (RNP) complexes. Lipofectamine Stem reagent is compatible with a variety of media systems, including feeder-free, helping support and simplify your stem cell culture workflow. Researchers can achieve up to 80% or better transfection efficiency in pluripotent stem cells (PSCs) and neural stem cells (NSCs), and up to 45% in mesenchymal stem cells (MSCs).
Transfecting stem cells without inhibiting cell viability and cell growth can be challenging due to the sensitivity of these cells. Lipofectamine Stem is a low toxicity reagent, requiring low amounts of nucleic acid, allowing your stem cells to stay healthy and continue proliferating without inducing differentiation. Lipofectamine Stem reagent is:
- Superior in performance—up to 3-fold higher efficiency across iPSCs, hESCs, NSCs, and MSCs compared to other reagents
- Versatile—one reagent for DNA, RNA, Cas9 protein, and co-transfection
- Gentle on cells—maintains cell proliferation without inducing differentiation
- Flexible—transfects cells in adherence or suspension—an alternative to electroporation
Achieve high efficiency gene editing in stem cells
Gene editing applications typically require the ability to co-deliver DNA, RNA, and/or protein into cells. Lipofectamine Stem Transfection Reagent is designed to co-deliver CRISPR/Cas9 complexes into human stem cells for high efficiency gene editing. This one reagent can be used to transfect large DNA plasmids such as all-in-one CRISPR/Cas9 vectors and co-deliver a range of RNAs, such as large Cas9 mRNA with tracr/crRNA complexes or single guide (sg) RNA, and protein such as Cas9 protein combined with tracr/crRNA or sgRNA.
A low-toxicity alternative to electroporation
Transfecting stem cells via electroporation methods can be challenging and cumbersome. Electroporation protocols recommend single cell suspension culture to ensure maximum efficiency and minimal toxicity. Using the reverse transfection protocol, the Lipofectamine Stem Transfection Reagent proves to be an effective and less toxic alternative to electroporation for stem cells. It requires low amounts of nucleic acid to achieve maximum transfection efficiency, on par with electroporation, while supporting continued proliferation without inducing differentiation.
Specifications
Specifications
| Classification | Animal Origin-Free |
| Content And Storage | Store in refrigerator (2°C to 8°C) and protect from light |
| Cell Type | Stem Cells |
| Sample Type | Cells |
| Transfection Technique | Lipid-based Transfection |
| For Use With (Application) | Transfection |
| Product Line | Lipofectamine |
| Product Type | Transfection Reagent |
| Quantity | 1.5 mL |
| Serum Compatible | No |
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Frequently Asked Questions (FAQs)
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.
For Research Use Only. Not for use in diagnostic procedures.