Lipofectamine Stem Transfection Reagent

Achieve up to 80% transfection efficiency in pluripotent stem cells (PSCs) and neural stem cells (NSCs), and up to 45% in mesenchymal stem cells (MSCs).

Explore the benefits of Lipofectamine Stem Transfection Reagent:

• Versatile—co-delivers DNA (up to 11 kb), RNA, and Cas9 protein complexes
• Gentle on cells—delivers low toxicity and continues cell proliferation without inducing differentiation
• Flexible—transfects adherent and suspension cells, offering a simple alternative to electroporation

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Although stem cell transfection is a a key technique for many researchers, the sensitivity of stem cells has made them difficult to transfect efficiently using reagent-based protocols. Designed for dramatically improved transfection efficiency, Invitrogen Lipofectamine Stem Transfection Reagent offers a simple, robust, and reproducible method for delivering DNA, RNA, and RNP, such as Cas9/gRNA complexes into a wide range of stem cells, including pluripotent stem cells, neural stem cells, and mesenchymal stem cells (Figure 2). This Lipofectamine transfection reagent family helps ensure high-efficiency transfection while maintaining maximum cell viability and growth in an undifferentiated state.

Research applications such as gene editing, gene expression, and directed differentiation depend on the effective, efficient delivery of DNA, RNA and/or Cas9 protein complexes into stem cells. Experiments involving large constructs have typically been carried out using electroporation, as reagent-based transfection yielded poor results. Lipofectamine Stem reagent offers expanded capabilities for stem cell transfection, making it possible to deliver DNA plasmids in sizes up to 11 kb.

For gene editing, Lipofectamine Stem reagent allows the co-transfection of Cas9 protein complexed with guide RNAs along with single-stranded DNA for homology directed repair. In addition, Lipofectamine Stem reagent can be used for the transfection of mRNA for gene expression or directed differentiation studies using stem cells. All of these attributes make this versatile reagent an ideal addition to your stem cell research.

When transfecting stem cells, it is important that culture conditions support their continued proliferation in an undifferentiated state. While introduction of exogenous constructs can have intended and unintended consequences, Lipofectamine Stem Transfection Reagent does not adversely affect proliferation (Figure 5).

There are many choices of culture systems for pluripotent stem cells including feeder-based and feeder-free systems, which include using a variety of matrices. Lipofectamine Stem reagent can be used in conjunction with a variety of culture systems. Below are some tips to maximize transfection efficiency in your culture system of choice:

1. For culture conditions where PSCs are passaged as clusters of cells and grown as large colonies on feeders or on Geltrex matrix, 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. This option can be used with monolayer culture systems as well to ensure even, high transfection efficiency or as an alternative to electroporation.
2. When transfecting adherent cultures growing on Geltrex or Matrigel matrices, it is important to plate small clusters or single pluripotent stem cells at an optimal density of 30–60% to allow them room to continue to expand during a transfection time of 24 to 48 hours.
3. When transfecting adherent cultures, growing cells on vitronectin or laminin—which allow the cells to spread out in more of a monolayer—can maximize transfection.
4. While Lipofectamine Stem reagent is compatible with most culture systems, components in some stem cell media can interfere with liposomal-based transfection. In those cases, using Opti-MEM medium for an initial transfection period of 1–4 hours followed by the addition of your medium of choice can minimize this effect while still supporting continued expansion. Refer to the Lipofectamine Stem Transfection Reagent user guide for specific details.

In addition to the Lipofectamine Stem Transfection Reagent user guide, please see stem cell-specific protocols that will help guide you to high efficiency transfection with low toxicity in individual stem cell types (ESC, NSC, or MSC) using your media of choice. These protocols cover growing cells, culture conditions, and passaging through to successful transfection results.