cat no | ioEA1003S Early Access
ioGABAergic Neurons are human induced pluripotent stem cell (iPSC)-derived GABAergic neurons, precision reprogrammed using opti-ox™ technology. Within 12 days post-revival, ioGABAergic Neurons form a pure, defined population that is ready for experimentation, expressing (>95%) classical marker genes including GAD1, GAD2, VGAT, DLX1 and DLX2. Calcium imaging has been used to demonstrate that ioGABAergic Neurons display spontaneous activity.
ioGABAergic Neurons provide an easy to use, high-quality human model for the study of neurological circuitry, neurodevelopmental and neurodegenerative disease research and drug development.
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Highly Pure
>95% of cells express key GABAergic markers within 12 days post-thaw, confirmed by single cell RNA sequencing.
Consistent
Get reproducible results from every vial with high lot-to-lot consistency, confirmed with bulk RNA sequencing.
Co-culture compatible
Suitable for co-culture and tri-culture studies with astrocytes and glutamatergic neurons.
Starting material
Human iPSC line
Karyotype
Normal (46, XY)
Seeding compatibility
6, 12, 24 & 96 well plates
Shipping info
Dry ice
Donor
Caucasian adult male (skin fibroblast)
Vial size
Small: >3 x 10⁶ viable cells
Quality control
Sterility, protein expression (ICC) and gene expression (RT-qPCR)
Differentiation method
opti-ox cellular reprogramming
Recommended seeding density
150,000 cells/cm²
User storage
LN2 or -150°C
Format
Cryopreserved cells
Product use
ioCells are for research use only
Applications
Disease research
Co-culture studies
Calcium imaging
Transcriptome analysis
MEA analysis
ioGABAergic Neurons generated by transcription factor-driven reprogramming of iPSCs using opti-ox technology
Time-lapse video capturing the rapid and homogeneous neuronal phenotype acquisition upon thawing of cryopreserved ioGABAergic Neurons. 12 day time course.
ioGABAergic Neurons express GABAergic neuron-specific markers
Immunofluorescent staining on day 10 post-revival demonstrates that ioGABAergic Neurons are positive for GABA (yellow), the pan-neuronal marker MAP2 (red), and the DAPI counterstain (blue).
ioGABAergic Neurons show mature neuronal networks by day 12
Upon reprogramming, ioGABAergic Neurons show rapid morphological changes, with neurons identified by day 7 post-revival. Mature neuronal networks are observed by day 12. Images show day 1 to 12 post-thawing; 10X magnification; scale bar: 200 μm.
Single-cell RNA sequencing shows ioGABAergic Neurons form a pure population (>95%) of mature GABAergic neurons
Single cell RNA sequencing analysis was performed with ioGABAergic Neurons at three specific timepoints (day 0, 7 and 14). By day 7, the population has a distinct expression profile indicating a pure population (>95%) of mature post-mitotic GABAergic neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells.
Single cell RNA sequencing shows mature ioGABAergic Neurons express key GABAergic markers
Single cell RNA sequencing analysis was performed with ioGABAergic Neurons at three specific timepoints (day 0, 7 and 14). By day 7, the expression of key GABAergic marker genes (GAD1, GAD2, SLC32A1 (VGAT), DLX2, DLX5), together with the pan-neuronal marker MAP2, could be detected in post-mitotic GABAergic neurons. Gene expression was assessed by 10x Genomics scRNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells.
Single-cell RNA sequencing shows mature ioGABAergic Neurons show minimal expression of markers indicative of other neuronal lineages
Single cell RNA sequencing analysis was performed with ioGABAergic Neurons at three specific timepoints (day 0, 7 and 14). By day 14, the expression of markers of non-GABAergic neuronal lineages (TH - dopaminergic, TPH1 - serotonergic, CHAT - cholinergic, SLC17A6 (VGLUT2) - glutamatergic, SLC17A7 (VGLUT1) - glutamatergic, SLC18A3 (VACHT) - cholinergic) are largely absent. Gene expression was assessed by 10x Genomics scRNA-sequencing. Note, this data is from cells in continuous culture and not cryopreserved cells.
Single cell RNA sequencing indicates that ioGABAergic Neurons are of the SST sub-type
ioGABAergic Neurons are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended media. The protocol for the generation of these cells is a three-phase process: Induction, which is carried out at bit.bio (Phase 0), Stabilisation for 3 days (Phase 1), and Maintenance (Phase 2) during which the ioGABAergic Neurons mature. Phases 1 and 2 after revival of cells are carried out at the customer site.
Mark Kotter | CEO and founder | bit.bio
Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology | Stanford University
2023
Everlum Bio & bit.bio
2023
V1
bit.bio
2022
Dr Deepak Srivastava | King’s College London
Download this infographic to find out how the approach used to generate human iPSC-derived cells influences purity, batch consistency and protocol speed.