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ioGlutamatergic Neurons have been generated from human induced pluripotent stem cells (iPSC) by Neurogenin-2 (NGN2)1 driven opti-ox2 reprogramming. Human stem cells, within days, convert into mature and functional glutamatergic neurons providing a high-quality cellular model with simple protocols for the study of neurological activity and disease.
ioGlutamatergic Neurons generated by NGN2-driven reprogramming of iPSCs using opti-ox technology.
Seven-day time course video, courtesy of bit.bio, capturing the rapid morphological changes from iPSCs upon induction of NGN2 expression. The video shows early neurons reaching the post-mitotic stage by day four and starting to build a dense neural network.
ioGlutamatergic Neurons are characterized by immunocytochemistry, gene expression, and bulk RNAseq. ioGlutamatergic Neurons cultures consist mainly of glutamatergic neurons (>80%) characterized by homogenous expression of pan-neuronal proteins MAP2 and TUBB3, and glutamate transporter genes VGLUT1 and VGLUT2. The minor remaining fraction of the neuronal population express marker genes of cholinergic neurons.
Immunofluorescent staining on post-revival day 11. Data courtesy of bit.bio
Examples of MaxOne high-resolution multi-electrode array (MEA) recordings of ioGlutamatergic Neurons in BrainPhys? media. The activity maps show the percentage of active electrodes (A), firing rate (B), and spike amplitude (C). A time-dependent increase of spontaneous activity during neuronal maturation from two to three weeks post-revival3 is demonstrated in the results.
Although iPSC-derived neurons have promising applications in drug discovery platforms, their use within high throughput screening (HTS) has previously been limited by traditional ‘directed differentiation’ approaches, making it difficult to generate reliable, homogenous cell populations at scale. Human iPSC-derived ioGlutamatergic Neurons overcome these restrictions to provide a robust platform for drug discovery, showing good suitability for high-throughput screening in 384-well plates.
Ease of Use: Cells arrive programmed to rapidly mature upon revival with only one medium required in a two-step protocol.
Consistency: Batch-to-batch reproducibility and homogeneity create a stable human model for excitatory neuronal activity and disease.
Speed: Ready for experimentation as early as two days post revival and allow the formation of functional neuronal networks at 17 days.
Scalability: Industrial scale quantities allow the cells to be used in a range of applications, from research to screening purposes.
|ioGlutamatergic Neurons (Human iPSC-derived glutamatergic neurons)||ab259259|
|CNQX, AMPA / kainate antagonist||ab120017|
|Tetrodotoxin, Na+ channel blocker||ab120054|
|Y-27632 dihydrochloride, Rho kinase inhibitor||ab120129|
|Doxycycline hyclate, matrix metalloprotease inhibitor||ab141091|
|Cytarabine, Pyrimidine nucleoside analog||ab141924|
2. Pawlowski, M. et al. Inducible and Deterministic Forward Programming of Human Pluripotent Stem Cells into Neurons, Skeletal Myocytes, and Oligodendrocytes. Stem Cell Reports. 8(4), 803-812 (2017).?