Stem Cell Intel Issue 8
Each quarter, Stem Cell Intel will provide you with the latest product news, upcoming industry events and easy access to technical tools such as publications, protocols, FAQs and more. To receive Stem Cell Intel, please click the subscribe button below.
Stay on the leading edge of stem cell research
The third annual 24 Hours of Stem Cells virtual event is running concurrently around the world for 24 hours on December 3, 2015. Join us from around the world to:
- Access cutting-edge scientific presentations in various formats, including live keynote speakers, live panel discussions and on-demand presentations
- Participate in unique educational opportunities, including online training and certifications
- Expand your global research network with opportunities to interact with scientists from across the globe
Experience all of this virtually, from the comfort of anywhere. Registration is free.
Optimized transfection reagent for CRISPR-Cas9 protein delivery
Invitrogen Lipofectamine CRISPRMAX is the first optimized lipid nanoparticle transfection reagent for CRISPR-Cas9 protein delivery, providing the cleavage efficiency of electroporation with the simplicity and scalability of a reagent. CRISPRMAX is an ideal alternative to electroporation as it is gentler on cells and more cost effective overall. Compared to other CRISPR reagents, it effectively delivers protein, allowing for fewer off-target effects. Deliver our superior GeneArt Platinum Cas9 Nuclease as well as other CRISPR-Cas9 proteins with a reagent that provides:
- Demonstrated cleavage efficiency in over 20 cell types, including iPSC, mESC, N2A, CHO, A549, HCT116, HeLa, HEK293 and several others
- Gentle transfection (low toxicity), which means less cells are needed to initiate your experiment
- The lowest cost, whether cost-per-reaction or initial investment
- An ideal delivery solution for high-throughput experiments
Recombinant Human Laminin-521 improves feeder-free reprogramming efficiency
Recently launched Gibco Recombinant Human Laminin-521 (rhLaminin-521) offers a significant improvement in efficiency when reprogramming PSCs under feeder-free conditions. (A) Representative alkaline phosphatase imaging. (B) Bar graph depicting number of alkaline phosphatase positive colonies achieved for the rhVTN-N and rhLaminin-521 conditions.
Human dermal fibroblasts, neonatal were expanded in Medium 106 with LSGS. HDFn were then reprogrammed in KSR-based medium using the CytoTune-iPS 2.0 Sendai Reprogramming Kit at an MOI of 5:5:3. On day 7 post transduction, newly reprogrammed fibroblasts were passaged onto rhLaminin-521 or VTN-N (rhVTN-N) matrices and were fed daily with Essential 8 Medium. On day 21 post transduction, the number of alkaline phosphatase colonies was determined per condition (n=3 per condition).
Webinar: Eliminating daily feeding in a feeder-free, xeno-free PSC culture system
Learn how to eliminate daily feeding from your PSC culture with Gibco Essential 8 Flex Medium. In this free, on-demand webinar we introduce Essential 8 Flex Medium and its many benefits, including:
- Extension of the activity of key heat-sensitive components including FGF 2
- Elimination of the need for daily feeding
- Allowance for routine weekend-free maintenance and expansion of PSCs without compromising culture performance
FAQ: Essential 8 Flex Medium
Thinking about optimizing your PSC culture workflow with the new Essential 8 Flex Medium? Have questions? We have the resource for you. Our Essential 8 Flex Medium FAQ document addresses all of your questions and concerns.
Figure 1. Images of H9 cultures 4 days following seeding. Confluence measurements were performed using automated image analysis and values indicated in upper left corner of each panel. For this set of H9 cultures, optimal confluency was about 60–80%.
Figure 2. Cardiomyocyte enrichment. Cardiomyocyte cultures were exposed to a glucose-free, lactate-containing medium (red bars) or to normal cardiomyocyte maintenance medium (blue bars). Noticeable enrichment was observed after 7 days via cytometric analysis.
Protocol: PSC Cardiomyocyte Differentiation Kit
Due to inherent variability between PSC lines, obtaining consistent and efficient cardiomyocyte differentiation across experiments can be challenging. New guidelines are now available to maximize your success using the Gibco™ PSC Cardiomyocyte Differentiation Kit:
- Better results for each new PSC line—A critical variable for the generation of robust cardiomyocyte culture is the relative confluence at the onset of differentiation (Figure 1). We strongly recommend performing a range-finding study when using a PSC line for the first time. Useful guidance for performing these studies can now be found in the product manual.
- Clean up your differentiated cultures—By switching from glucose to lactate as an energy source after differentiation, non-cardiomyocytes will not survive, leading to an enriched population of cardiomyocytes (Figure 2).
Access optimized protocol
For Research Use Only. Not for use in diagnostic procedures.