Gain a deeper understanding of cellular complexities and tease apart variations due to age, gender, and donor variability using neurons derived from human iPS cells.
Differentiation kit workflow
Representative phase contrast images of Quick-Endothelium™ Vascular – mRNA Kit cell cultures on days 1-5 post-differentiation (scale bar = 100 μm).
iPSC-derived cells workflow
Representative phase contrast images of Quick-Endothelium™ Vascular - Human iPSC-derived vascular endothelial cell cultures on days 1, 2, and 3 post-thaw (scale bar = 100 μm).
Immunofluorescent staining of Quick-Endothelium™ Vascular - mRNA Kit cell cultures shows typical vascular endothelium morphology and expression of CD31 on day 5 post-differentiation (scale bar = 100 μm). Staining conditions: Anti-CD31 primary antibody (Biolegend, Catalog Number: 303102, 1:50 dilution) in combination with a secondary antibody (Invitrogen, Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594, Catalog Number:A11032, 1:500 dilution) were applied. Nuclei were counterstained with Hoechst 33342.
Immunofluorescent staining of Quick-Endothelium™ Vascular - mRNA Kit cell cultures shows typical vascular endothelium morphology and expression of endothelial markers CD31 and vWF as well as the tight junction marker Claudin-5 on Day 6 (scale bar = 100 μm). The expression levels of each marker protein were comparable to those of HUVECs.
Images courtesy of Ricoh.
Comparison of lumen forming ability of Quick-Endothelium™ Vascular cells and HUVECs. 5×105 Quick-Endothelium™ Vascular cell were plated on Matrigel-coated 24-well plates and incubated for 24 hours for comparison with HUVECs seeded at a density of 1×105 cells. Quick-Endothelium™ Vascular cells formed a reticular structure similar to HUVECs and sheet-like areas were observed.
Images courtesy of Ricoh.
Comparison of cytokine reactivity (IL-6) in Quick-Endothelium™ Vascular cells and HUVECs. Cells were seeded onto 96-well plates at a density of 2×104cells/well (HUVEC) or 5×104cells/well (Quick-Endothelium™ Vascular cells) and incubated for 48h. IL-1β was added and cells were further incubated for 24h. Supernatant was collected and secretion of IL-6 was measured. Change in IL-6 was calculated relative to control unstimulated cells.
Data courtesy of Ricoh.
Comparison of cytokine reactivity (IL-8) in Quick-Endothelium™ Vascular cells and HUVECs. Cells were seeded onto 96-well plates at a density of 2×104cells/well (HUVEC) or 5×104cells/well (Quick-Endothelium™ Vascular cells) and incubated for 48h. LPS was added and cells were further incubated for 24h. Supernatant was collected and secretion of IL-8 was measured. Change in IL-8 was calculated relative to control unstimulated cells.
Data courtesy of Ricoh.
Our Quick-Endothelium™ Vascular - mRNA Kits allow researchers to quickly, easily, and efficiently differentiate their iPS or ES cell line of choice into vascular endothelial cells. The kits utilize synthetic messenger RNA (syn-mRNA) to deliver our proprietary cocktail of transcription factors that induce differentiation without leaving a genetic footprint.
Quick-Endothelium™ Vascular - mRNA Kits are available by request. Please inquire via email!
Elixirgen Scientific’s Quick-Endothelium™ Vascular - Human iPSC-derived Endothelial Cells are created using human iPS cells made available through a license from The California Institute for Regenerative Medicine (CIRM).
The CIRM iPS cell repository includes over 1,500 iPS cell lines, so please contact us if you would like vascular endothelial cells derived from a specific donor cell line. Every lot of iPSCs is differentiated into vascular endothelial cells using our proprietary transcription factor-based technology that is “footprint-free” and delivers functional, high-quality cells.
Elixirgen Scientific’s Quick-Endothelium™ Vascular - Human iPSC-derived Endothelial Cells are available by request. Please inquire via email!
User guides are available by request. Please inquire via email!
No. The use of differentiated cells or kits provided by Elixirgen Scientific does not require any additional license from other parties for any type of use, except for use in humans or for therapeutic or diagnostic use. Check out our license statement in Resource section for further detail.
Off the shelf, we offer small (>1 million viable cryopreserved cells) and large (5x >1 million viable cryopreserved cells) sizes. If you are looking for a larger quantity, please contact us for pricing information.
Both delivery methods provide robust “footprint-free,” non-integrating expression of the transcription factors, although each kit uses a slightly different workflow. Based on our data, the differentiation efficiency between an mRNA-based and a SeV-based kit is similar. For example, the percentages of TUBB3+ neurons differentiated using Quick-Neuron™ Cholinergic mRNA- and SeV-based kits were 93% and 88%, respectively.
We recommend the mRNA-based neuron differentiation kits for labs with experience culturing iPSCs/ESCs, as the workflow involves slightly more manipulation of fragile differentiating cells than the SeV-based kits. The SeV-based kits are better for labs with little experience culturing iPSCs/ESCs, although they must be used with BSL-2 safety measures and may require prior approval from your Institutional Biosafety Committee (IBC).
Off the shelf, we offer small and large iPSC neuronal differentiation kits. The small size is best suited for single cell, imaging analyses, and pilot studies while the large size is best for applications such as high throughput screening and RNA sequencing.
