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Human iPSC-derived GABAergic neurons

ioRD003X | In development

ioGABAergic Neurons have been reprogrammed from human induced pluripotent stem cells (iPSCs) using our precise reprogramming technology: opti-ox™1 (optimised inducible overexpression). Human stem cells, within days, convert into consistent, functional inhibitory GABAergic neurons providing a high-quality human model for the study of neurological connectivity and disorders.

ioGABAergic Neurons, part of our ioCells portfolio, consist of GABAergic neurons (>95%) characterised by the expression of the classical marker genes including GAD1, GAD2, VGAT, DLX1 and DLX2. Single cell RNA sequencing analysis shows that ioGABAergic Neurons have characteristics of the vasoactive intestinal polypeptide (VIP) subtype of GABAergic neurons and rapidly form neuronal networks with spontaneous neuronal activity by day 20 post-revivial. 

Ready-to-culture cells are suitable as models for research in cell-type specific biology,  target validation and drug screening in pharmaceutical R&D.

¹ Pawlowski et al., Stem Cell Report 2017

Product specifications

Advantages

Ready for experimentation within days
Highly defined 
In depth single cell characterisation
Easy culturing

Applications

– Academic research
– Co-cultures
– High-content screening
– Axonal development studies
– Single cell RNA sequencing**

For more information email info@bit.bio

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 ioGABAergic Neurons generated by transcription factor driven reprogramming of iPSCs using opti-ox™ technology

Video capturing the rapid morphological changes from human iPSCs upon transcription factor expression using opti-ox™ cellular reprogramming. 5 day time course.

 
ioGABAergic Neurons express GABAergic neuron-specific markers

VGAT image 374px

VGAT

GABA image 374px

GABA

MAP2 image 374px

MAP2

Merge image 374px

Merge

 Immunofluorescent staining on post-revival Day 14 demonstrates that ioGABAergic Neurons are positive for VGAT (green) and GABA (red) as well as for the pan-neuronal marker MAP2 (cyan) and DAPI counterstain (blue).

ioGABAergic Neurons show mature neuronal networks by Day 11

Day 1 picture

Day 1

Day 4 picture

Day 4

Day 7 picture

Day 7

Day 11 picture

Day 11

Rapid morphological changes in the cells upon reprogramming, with neurons identified by Day 7 post-revival. Mature neuronal networks are observed by Day 11. Day 1 to 11 post-thawing; 100X magnification; scale bar: 200μm.

 
Highly-defined and characterised by single cell RNA sequencing

ioGABAergic Neuron - Single cell RNA sequencing

 

Expression of a specific transcription factor combination efficiently converts iPSCs into GABAergic neurons. From this continuous culture the post-mitotic neurons, at Day 14, express key marker genes of GABAergic neurons (DLX1, GAD1, GAD2, VGAT) together with pan-neuronal marker MAP2 (left panel). Gene expression indicative of other neuronal lineages (glutamatergic, dopaminergic, cholinergic or serotonergic) are largely absent (right panel). Gene expression assessed by 10x Genomics scRNA-sequencing.

ioGABAergic Neurons show characteristics of the vasoactive intestinal polypeptide (VIP) subtype

Bioinformatic analysis of GABAergic subtype identity-1

 

Bioinformatic analysis of GABAergic subtype identity from single cell RNA-seq data. Anchoring our single cell RNA-seq data of the iPSC-derived GABAergic neurons on the Allen Brain single cell map of the human cortex demonstrates that the ioGABAergic Neurons (red cluster) closely match with the VIP subtype of GABAergic neurons (purple cluster).

Cells arrive ready to plate

bit.bio-iogabaergic-neurons_vial_v3_retina

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 with Doxycycline (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.

 Product specifications

Starting material
human iPSC line

Donor
caucasian adult male
(skin fibroblast)

Differentiation method
opti-oxTM cellular reprogramming

Karyotype
contact bit.bio

Vial size
contact bit.bio

Recommended seeding density
contact bit.bio

Seeding compatibility
contact bit.bio

Quality control
Sterility, protein expression (IF) and gene expression (RT-qPCR)

User storage
LN2 or -150°C
 

Shipping info
Dry ice

Product use
These cells are for research use only


Supporting documentation


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