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cat no | io1110 Early Access
ioAstrocytes 2.0
Human iPSC-derived astrocytes
- Cryopreserved human iPSC-derived cells powered by opti-ox™, assay-ready in days using serum free culture media
- Optimised for multi-cellular in vitro CNS models, compatible with MEA electrophysiology applications
- Rapidly displaying key functional capabilities, including supporting robust neuronal network activity in MEA assays, phagocytosis, and cytokine secretion
Human iPSC-derived astrocytes
bit.bio ioAstrocytes 2.0 adopt classic stellate morphology and express key astrocyte markers. Representative immunocytochemistry staining of vimentin (magenta), with DAPI as nuclear counterstain (blue).
Microelectrode array (MEA) functional characterisation of co-cultured ioGlutamatergic Neurons and ioAstrocytes 2.0
Representative MEA raster plots captured using the Axion BioSystems Maestro MEA platform evaluating co-cultures of 120K ioGlutamatergic Neurons and ioAstrocytes 2.0 (3:1 seeding ratio) over an extended maturation timeline (DIV29 to DIV50).
Each panel depicts a 5-minute continuous electrical activity recording. At early time points (DIV29–DIV34), cultures exhibit predominantly localised asynchronous neuronal firing. By DIV36–DIV41, co-cultures establish mature, highly synchronised functional networks, characterised by the emergence of organised network bursting (highlighted by orange vertical overlay markers).
Network synchrony is maintained through DIV50.
Advanced tri-culture of ioGlutamatergic Neurons, ioAstrocytes 2.0, and ioMicroglia
While neuronal-astrocyte co-cultures capture essential structural and metabolic synergy, the addition of microglia, the resident myeloid cells of the CNS, is critical for accurately modelling neuroinflammatory pathways, synaptic pruning, and homeostatic maintenance.
This representative immunocytochemistry image demonstrates a successful tri-culture established at a 2:1:2 ratio (neurons:astrocytes:microglia). Using an optimised bit.bio protocol for 24-well plates, the tri-culture can be robustly maintained for up to 7 days. Staining with cell type-specific markers confirms the presence and integration of all three populations, shown alongside single-channel panels: MAP2-positive ioGlutamatergic Neurons (red), S100B-positive ioAstrocytes 2.0 (cyan), and IBA1-positive ioMicroglia (yellow); with DAPI as a nuclear counterstain (blue). Scale bars: 200 µm.
Robust co-culture protocol of ioGlutamatergic Neurons and ioAstrocytes 2.0
By combining neuronal and glial cells, this in vitro co-culture more accurately recapitulates the complex, heterogeneous in vivo environment of the human brain, maximising physiological relevance for research.
This representative immunocytochemistry image demonstrates day 17 of a successful co-culture established at a 1:1 ratio using an optimised bit.bio protocol in 24-well plates. Staining with cell type-specific markers confirms the presence and integration of the two populations: MAP2-positive ioGlutamatergic Neurons (red), S100B-positive ioAstrocytes 2.0 (cyan), along with a nuclear counterstain (DAPI, blue). Scale bars: 200 µm; 20x magnification.
This protocol is adaptable and supports direct substitution with products of the same cell types, such as ioGlutamatergic Neurons disease models and GFP ioGlutamatergic Neurons.
ioAstrocytes 2.0 secrete cytokines in response to stimulation
Cells were treated with three distinct pro-inflammatory cocktails or a vehicle control at days 7 and 14 post-thaw. Following a 24-hour incubation, conditioned media was collected and cytokine concentrations were quantified via MSD multiplex immunoassay. Pro-inflammatory treatments broadly upregulated cytokine secretion relative to vehicle controls, eliciting expected neuroinflammatory responses, including the robust secretion of Interleukin-6 (IL-6).
Note: Exogenous IFNγ and IL-1β present in two of the stimulation cocktails preclude accurate measurement of their endogenous secretion in those specific conditions.
ioAstrocytes 2.0 secrete cytokines in response to stimulation
Cells were treated with three distinct pro-inflammatory cocktails or a vehicle control at days 7 and 14 post-thaw. Following a 24-hour incubation, conditioned media was collected and cytokine concentrations were quantified via MSD multiplex immunoassay. Pro-inflammatory treatments broadly upregulated cytokine secretion relative to vehicle controls, eliciting expected neuroinflammatory responses, including the robust secretion of Interleukin-6 (IL-6).
Note: Exogenous IFNγ and IL-1β present in two of the stimulation cocktails preclude accurate measurement of their endogenous secretion in those specific conditions.
Immunocytochemistry shows protein expression of key astrocyte markers
ioAstrocytes 2.0 express the key astrocyte markers S100B and SOX9 at days 7 and 14 post-thaw. DAPI was used as a nuclear stain.
S100B is a multifaceted protein primarily found in astrocytes playing a key role in activation, neuroprotection, calcium homeostasis and astrocyte-neuron communication. SOX9 is critical for the differentiation of astrocytes.
Immunocytochemistry shows protein expression of vimentin
ioAstrocytes 2.0 express the key astrocyte marker vimentin at days 7 and 14 post-thaw. DAPI was used as a nuclear stain.
Vimentin is a cytoskeletal protein enriched in astrocytes.
RT-qPCR shows gene expression of key astrocyte markers
RT-qPCR data showing expression of key astrocyte markers EAAT1, SOX9, S100B and Vimentin (VIM) at four different timepoints (iPSC, D1, D7 and D14). ioAstrocytes 2.0 show expression of key markers from as early as day 1 post-thaw. Pluripotency markers POU5F1 (OCT4) and NANOG are downregulated.
The SLC1A3 gene encodes Excitatory Amino Acid Transporter 1 (EAAT1), a protein that plays a crucial role in regulating glutamate neurotransmission, maintaining neuronal health, and protecting against excitotoxicity.
View the step-by-step RNA extraction and RT-qPCR protocol used to generate this 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.
Benchtop benefits
Co-culture ready
ioAstrocytes 2.0 support functional neuronal networks within co-culture settings, enabling in-vitro modelling of complex CNS biology.
Functional
Display key phagocytic and cytokine secretion functions and demonstrate a direct influence on neuronal network activity through validated MEA assays.
Easy to use
Cells arrive programmed to mature rapidly upon revival and are maintained using serum-free components.
Cells arrive ready to plate
ioAstrocytes 2.0 are delivered in a cryopreserved format and are programmed to mature rapidly upon revival in the recommended serum-free media. The protocol for the generation of these cells is a two-phase process: 1. Stabilisation for 5 days. 2. From day 5 onwards, maintenance of cells according to the protocol and recommended media for the duration of assay requirements.
What scientists say about ioAstrocytes
Jeremy Krohn
PhD Candidate | DZNE / Charité University of Medicine
"We used bit.bio human ioAstrocytes in our experiments to compare with - and step-wise replace - animal derived cells. The human cells were straightforward to establish, showed high reproducibility, and performed well in calcium imaging experiment."
Rapid gain of functional activity
ioAstrocytes 2.0 support robust neuronal co-culture network activity
Representative MEA raster plots captured using the Axion BioSystems Maestro MEA platform evaluating co-cultures of 120K ioGlutamatergic Neurons and ioAstrocytes 2.0 (3:1 seeding ratio) over an extended maturation timeline (DIV29 to DIV50).
Each panel depicts a 5-minute continuous electrical activity recording. At early time points (DIV29–DIV34), cultures exhibit predominantly localised asynchronous neuronal firing. By DIV36–DIV41, co-cultures establish mature, highly synchronised functional networks, characterised by the emergence of organised network bursting (highlighted by orange vertical overlay markers). Network synchrony is maintained through DIV50.
Phagocytosis of pHrodo Red S. aureus Bioparticles by ioAstrocytes 2.0
Incucyte® video showing the ability of ioAstrocytes 2.0 to phagocytose pHrodo™ Red S. aureus Bioparticles™.
The Bioparticles were added to cultures of ioAstrocytes 2.0 at 7 days post-thaw. Initially the particles are located outside the cells in the neutral pH of the medium and are non-fluorescent. When phagocytosed, the particles are exposed to the acidic environment of intracellular organelles and fluoresce bright red.
The video shows an increase of red fluorescent particles accumulating within the cells over a time course of 48 hours, demonstrating that ioAstrocytes 2.0 have the capacity to phagocytose.
Pro-inflammatory stimuli induce cytokine secretion in ioAstrocytes 2.0
Click on the tabs to explore the data.
Cells were treated with three distinct pro-inflammatory cocktails or a vehicle control at days 7 and 14 post-thaw. Following a 24-hour incubation, conditioned media was collected and cytokine concentrations were quantified via MSD multiplex immunoassay. Pro-inflammatory treatments broadly upregulated cytokine secretion relative to vehicle controls, eliciting expected neuroinflammatory responses, including the robust secretion of Interleukin-6 (IL-6).
Note: Exogenous IFNγ and IL-1β present in two of the stimulation cocktails preclude accurate measurement of their endogenous secretion in those specific conditions.
Highly characterised and defined
Immunocytochemistry shows protein expression of key astrocyte markers
Click on the tabs to explore the data.
ioAstrocytes 2.0 express the key astrocyte markers S100B, SOX9 (A) and Vimentin (B) at days 7 and 14 post-thaw. DAPI was used as a nuclear stain.
S100B is a multifaceted protein primarily found in astrocytes playing a key role in activation, neuroprotection, calcium homeostasis and astrocyte-neuron communication. SOX9 is critical for the differentiation of astrocytes. Vimentin is a cytoskeletal protein enriched in astrocytes.
RT-qPCR shows gene expression of key astrocyte markers
RT-qPCR data showing expression of key astrocyte markers EAAT1, SOX9, S100B and Vimentin (VIM) at four different timepoints (iPSC, D1, D7 and D14). ioAstrocytes 2.0 show expression of key markers from as early as day 1 post-thaw. Pluripotency markers POU5F1 (OCT4) and NANOG are downregulated.
The SLC1A3 gene encodes Excitatory Amino Acid Transporter 1 (EAAT1), a protein that plays a crucial role in regulating glutamate neurotransmission, maintaining neuronal health, and protecting against excitotoxicity.
View the step-by-step RNA extraction and RT-qPCR protocol used to generate this data.
Functional MEA co-culture
ioGlutamatergic Neuron–ioAstrocyte 2.0 co-cultures form mature, synchronised neuronal networks over time.
Representative MEA raster plots generated using the Axion BioSystems Maestro MEA platform show electrical activity in co-cultures of 120K ioGlutamatergic Neurons and ioAstrocytes 2.0, seeded at a 3:1 ratio, across an extended maturation period (DIV29–DIV50). Each panel represents a 5-minute continuous recording.
At early time points (DIV29–DIV34), cultures display predominantly localised, asynchronous neuronal firing. From DIV36–DIV41, co-cultures develop mature, highly synchronised functional networks, characterised by organised network bursting (orange vertical markers), with network synchrony maintained through DIV50.
In-vitro CNS co-culture model
By combining neuronal and glial cells, this in vitro co-culture more accurately recapitulates the complex, heterogeneous in vivo environment of the human brain, maximising physiological relevance for research.
This representative immunocytochemistry image demonstrates day 17 of a successful co-culture established at a 1:1 ratio using an optimised bit.bio protocol in 24-well plates. Staining with cell type-specific markers confirms the presence and integration of the two populations: MAP2-positive ioGlutamatergic Neurons (red), S100B-positive ioAstrocytes 2.0 (cyan), along with a nuclear counterstain (DAPI, blue). Scale bars: 200 µm; 20x magnification.
This protocol is adaptable and supports direct substitution with products of the same cell types, such as ioGlutamatergic Neurons disease models and GFP ioGlutamatergic Neurons.
In vitro CNS tri-culture model
Advanced tri-culture of ioGlutamatergic Neurons, ioAstrocytes 2.0 and ioMicroglia
While neuronal-astrocyte co-cultures capture essential structural and metabolic synergy, the addition of microglia, the resident myeloid cells of the CNS, is critical for accurate modelling of neuroinflammatory pathways, synaptic pruning and homeostatic maintenance.
This representative immunocytochemistry image demonstrates a successful tri-culture established at a 2:1:2 ratio (neurons:astrocytes:microglia). Using an optimised bit.bio protocol for 24-well plates, the tri-culture can be maintained for up to 7 days. Staining with cell type-specific markers confirms the presence and integration of all three populations, shown alongside single-channel panels: MAP2-positive ioGlutamatergic Neurons (red), S100B-positive ioAstrocytes 2.0 (cyan), and IBA1-positive ioMicroglia (yellow); with DAPI as a nuclear counterstain (blue). Scale bars: 200 µm.
This protocol is adaptable and supports direct substitution with products of the same cell types, such as ioGlutamatergic Neurons disease models, ioMicroglia disease models and GFP reporter cells.
ioAstrocytes_2.0_Hero_image_VIM_DAPI.webp?width=600&name=(UPDATED)ioAstrocytes_2.0_Hero_image_VIM_DAPI.webp&format=webp&quality=80)
Expand your research
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Build neurodegeneration disease models
Model neurodegeneration in vitro with astrocyte-neuron co-cultures
Expand your research
Build neurodegeneration disease models
Model neurodegeneration in vitro with astrocyte-neuron co-cultures
Access 20 neuronal disease models with a single co-culture protocol.
View the co-culture protocol
Explore ioGlutamatergic Neuron Disease Models
Generate functional in vitro models of the CNS
Highly pure human iPSC-derived cells to build complex multi-cell cultures
Expand your research
Generate functional in vitro models of the CNS
Highly pure human iPSC-derived cells to build complex multi-cell cultures
ioAstrocytes are highly pure, defined and consistent, a great companion for your CNS model.
Combine them with other neuronal and glia ioCells to generate complex multi-cell cultures.
ioGlutamatergic Neurons
ioMicroglia
ioOligodendrocyte-like cells
Study neuroinflammation
Assess cell damage, microglia activation and cytokine release
Expand your research
Study neuroinflammation
Assess cell damage, microglia activation and cytokine release
Interested in optimising your neuroinflammation studies?
Move into a multi-cellular model using different cell types:
ioMicroglia Male
ioGlutamatergic Neurons
ioOligodendrocyte-like cells
Tailor experimental conditions to answer your scientific question by including ioDisease Model Cells with disease-related mutations.
Study disease-related mutations
Make true comparisons with a genetically-matched control
Expand your research
Study disease-related mutations
Make true comparisons with a genetically-matched control
Build your custom disease model into the ioAstrocytes wild type. Use the genetically matched control in your experiments to make true comparisons in your data, being confident that even subtle variances in your data are attributable to your mutation of interest.
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