cat no | io1115
ioGlutamatergic Neurons MAPT S305N/S305N
Human iPSC-derived neuronal 4R tau model
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Cryopreserved human iPSC-derived cells powered by opti-ox that are ready for experiments in days
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Functional excitatory neurons engineered with the MAPT S305N mutation for FTD, Alzheimer's disease and tauopathy research
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Nearly equimolar ratio of 4R:3R MAPT demonstrated by RT-qPCR and Western blotting.
Human iPSC-derived neuronal 4R tau model
Expression of 4R tau in glutamatergic neurons carrying a S305N homozygous mutation in the MAPT gene.
ioGlutamatergic Neurons Media Kit
Cell culture media kit for the culture of ioGlutamatergic Neurons up to 14 days post-thaw
Adult-like 4R:3R tau isoform expression
Cells carrying the S305N mutation express nearly equimolar 4R:3R MAPT isoforms
MAPT isoform expression analysis via RT-qPCR, quantifying total, 3R and 4R MAPT expression in neuronal cultures at days 11, 25 and 32.
RNA (100 ng) was reverse-transcribed and amplified using TaqMan Fast Advanced Master Mix (Thermo Fisher Scientific) and isoform-specific probes (total MAPT: Hs00902194_m1; 3R MAPT: Hs00902192_m1, 4R MAPT: Hs00902312_m1). Expression levels were normalised to HMBS and the 4R:3R MAPT ratio was calculated.
Western blot confirms the presence of 4R tau protein in S305N mutant cells
Tau expression and phosphorylation profiles in MAPT mutant ioGlutamatergic Neurons
Western blot analysis of total tau, 4R tau, and pTau in WT vs. MAPT S305N/S305N (+P301S) ioGlutamatergic Neurons at Day 32. The Western blot for total tau (A) shows the presence of the 4R and 3R isoforms in all MAPT mutant neurons, indicated by the arrows, whereas the wild-type control (WT) shows only 3R tau.
Cells were lysed in RIPA buffer with protease inhibitors (A, B) or protease/phosphatase inhibitors (C); total protein was quantified via BCA assay. For total and 4R tau blots, lysates were pre-treated with lambda phosphatase (3h at 30°C).
Cell lysates and a tau ladder positive control were resolved on a 4–20% Mini-PROTEAN® TGX Stain-Free™ gel (120V, 75 min) and transferred to a PVDF membrane. For total tau 10 μg protein was loaded; for 4R and pTau 20 μg protein was loaded.
Membranes were blocked (10 min) and incubated overnight at 4°C with primary antibodies: Anti-Tau antibody [TAU-5] (Abcam #ab80579, 1:1000), Tau 4R (E7T4F) Rabbit Monoclonal Antibody (Cell Signalling #79327, 1:1000), Phospho-Tau (Ser202, Thr205) Monoclonal Antibody (AT8) (Thermo Fisher #MN1020, 1:1000), or for the loading control, Anti-GAPDH antibody [6C5] (Abcam #ab8245, 1:5000). Signal was detected via ECL following secondary antibody incubation.
Relative pTau band intensity (D) was quantified using ImageJ (arbitrary units).
Immunocytochemical analysis of 4R tau expression
4R Tau is absent in WT and most abundant in the S305N/S305N cells
Representative images of 4R tau expression in wild-type (WT) and MAPT S305N/S305N (+P301S) ioGlutamatergic Neurons at Day 31.
Cells were fixed in 4% PFA and stained for the neuronal marker MAP2 (Abcam #ab5392) and 4R tau (Cell Signalling #79327). Fluorescent signals were visualised using an Echo Revolve microscope (10x objective).
Phenotypic characterisation of a human iPSC-derived tri-culture using ioGlutamatergic Neurons, ioAstrocytes, and ioMicroglia
Using our fully optimised protocol, ioGlutamatergic Neurons (MAP2, red), ioMicroglia (IBA1, yellow) and ioAstrocytes (vimentin, cyan) were co-cultured to create a highly defined CNS model. High-resolution ICC analysis confirms the successful co-localisation and morphological health of three distinct cell types within a unified environment. By day 7, the protocol yields a highly consistent, integrated network suitable for complex cell modelling. DAPI (blue) highlights the total cell density and integrity of the culture. This protocol is compatible with derivative products of the three cell types, ensuring straightforward implementation across experimental workflows.
Efficient mRNA transfection into ioGlutamatergic Neurons
ioGlutamatergic Neurons are efficiently transfected and show sustained long-term expression of mRNA encoding GFP. ioGlutamatergic Neurons were imaged from day 1 post-thaw and throughout the experiment to assess transfection efficiency and evaluate potential cytotoxic effects of the transfection protocol. Day 1 images were captured prior to transfection on the same day.
Download the step-by-step protocol for lipid-based delivery of synthetic mRNA into ioGlutamatergic Neurons.
Lipid-based delivery of synthetic mRNA into ioGlutamatergic Neurons
ioGlutamatergic Neurons were transfected 24 hours post-thaw using Lipofectamine™ Stem Transfection Reagent. The transfection efficiency was evaluated by fluorescence imaging over 18 days after mRNA delivery, resulting in high transfection efficiency (close to 100%) and long-term sustained GFP expression.
Quantification of the GFP signal shows a decrease in GFP intensity over time, while the percentage of GFP-positive cells remains largely unchanged over time.
(A) The percentage of GFP-positive cells from two independent experiments.
(B) GFP intensity, quantified in successfully transfected cells from two independent experiments is quantified and normalised to day 2 (24 hours post-transfection).
Industry leading seeding density
The recommended minimum seeding density is 30,000 cells/cm2, compared to up to 250,000 cells/cm2 for other similar products on the market. One small vial can plate a minimum of 0.7 x 24-well plate, 1 x 96-well plate, or 1.5 x 384-well plates. This means every vial goes further, enabling more experimental conditions and more repeats, resulting in more confidence in the data.
Vial limit exceeded
A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@bit.bio.
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