cat no | ioEA1093 Early Access
Confidently investigate your phenotype of interest across multiple clones with our disease model clone panel. Detailed characterisation data (below) and bulk RNA sequencing data (upon request) help you select specific clones if required.
Co-culture ready
ioAstrocytes support functional neuronal networks within co-culture settings, enabling in-vitro modelling of complex CNS biology.
Functional
Display key phagocytic and cytokine secretion functions as well as a demonstrable influence on neuronal network activity.
Consistent
Get reproducible results from every vial with lot-to-lot consistency of highly characterised & defined human iPSC-derived cells.
ioAstrocytes 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: 1. Stabilisation for 2 days. 2. Pre-maintenance for an additional 6 days. 3. From day 8 onwards, maintenance of cells according to the protocol and recommended media for the duration of assay requirements.
This protocol describes a method of co-culturing ioAstrocytes with ioGlutamatergic Neurons and associated disease models to facilitate research into complex neuroglial interactions.
NanoLive video showing the ability of the ioAstrocytes to phagocytose pHrodo® Red S. aureus Bioparticles® for Incucyte®.
pHrodo® Red S. aureus Bioparticles® were added to cultures of ioAstrocytes at 15 days post-thaw. At the start of the video, the particles are located outside the cells and due to the neutral pH of the media are non-fluorescent, but when phagocytised they are exposed to the acidic environments of intracellular organelles and fluoresce bright red. The video shows an increase of red fluorescent particles accumulating within the cell over a time course of 24 hours, demonstrating that ioAstrocytes have the capability to phagocytose.
MSD multiplex immunoassay measuring whether ioAstrocytes are able to secrete a range of cytokines upon treatment with various proinflammatory stimuli. ioAstrocytes were treated with 3 different proinflammatory cocktails (T) or vehicle (V) at day 15 post-thaw and after 24 hours media samples were collected to measure the concentration of cytokines in the media.
The proinflammatory cocktails induce the secretion of most of the cytokines relative to the vehicle treated cells. Overall, the ioAstrocytes display the expected responses to the three distinct cocktails, including a strong response of Interleukin 6 (IL-6), known to be involved in neuroinflammation. Note that IFNg or IL-1b are present in two of the inflammatory cocktails and, therefore, the presence of these cytokines in the media will lead to high signals upon their detection and interfere with the measurements of their secreted forms.
Immunocytochemistry shows protein expression of key astrocyte markers
Click on the tabs to explore the data.
ioAstrocytes express at Day 15 and Day 22 the key astrocyte markers S100B, SOX9 (A) and Vimentin (B). 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 data showing expression of key astrocyte markers EAAT1, SOX9, S100B and Vimentin (VIM) at four different timepoints (iPSC & D8, D15, D22). ioAstrocytes show expression of key markers from as early as day 8. Pluripotency markers POU5F1 (OCT4) & NANOG) are downregulated.
The SLC1A3 gene codes for the EAAT1 protein (Excitatory Amino Acid Transporter 1). Playing crucial roles in the regulation of glutamate neurotransmission, maintaining neuronal health and protecting against excitotoxicity.
Bulk RNA-sequencing demonstrates high lot-to-lot consistency of ioAstrocytes
Bulk RNA-sequencing analysis was performed on three different lots of manufactured product at day 1 and day 22 post revival. Principal component analysis (PCA) represents the variance in gene expression between the three different lots of ioAstrocytes. This analysis shows lots clustering very closely (<0.5% differentially expressed genes) demonstrating high consistency at each given timepoint.
This lot-to-lot consistency of ioAstrocytes will help reduce experimental variation and increase the reproducibility of data. Colours represent the parental non-induced hiPSC cell line and the three lots of ioAstrocytes; shapes represent different timepoints.
Expression levels for specific genes of interest can be requested by contacting our team at technical@bit.bio.
Do more with every vial
The seeding density of our human iPSC-derived astrocytes has been optimised and validated to a recommended seeding density of 30,000 cells/cm2. This means scientists can do more with every vial and expand experimental design within budget without losing out on quality. Resulting in more experimental conditions, more repeats, and more confidence in the data. 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 plate.
This protocol describes a method of co-culturing ioAstrocytes with ioGlutamatergic Neurons and associated disease models to facilitate research into complex neuroglial interactions.
NanoLive video showing the ability of the ioAstrocytes to phagocytose pHrodo® Red S. aureus Bioparticles® for Incucyte®.
pHrodo® Red S. aureus Bioparticles® were added to cultures of ioAstrocytes at 15 days post-thaw. At the start of the video, the particles are located outside the cells and due to the neutral pH of the media are non-fluorescent, but when phagocytised they are exposed to the acidic environments of intracellular organelles and fluoresce bright red. The video shows an increase of red fluorescent particles accumulating within the cell over a time course of 24 hours, demonstrating that ioAstrocytes have the capability to phagocytose.
MSD multiplex immunoassay measuring whether ioAstrocytes are able to secrete a range of cytokines upon treatment with various proinflammatory stimuli. ioAstrocytes were treated with 3 different proinflammatory cocktails (T) or vehicle (V) at day 15 post-thaw and after 24 hours media samples were collected to measure the concentration of cytokines in the media.
The proinflammatory cocktails induce the secretion of most of the cytokines relative to the vehicle treated cells. Overall, the ioAstrocytes display the expected responses to the three distinct cocktails, including a strong response of Interleukin 6 (IL-6), known to be involved in neuroinflammation. Note that IFNg or IL-1b are present in two of the inflammatory cocktails and, therefore, the presence of these cytokines in the media will lead to high signals upon their detection and interfere with the measurements of their secreted forms.
Consistent. Defined. Scalable.
