Astrocytes are the most abundant glial cells found in the central nervous system and spinal cord, and are involved in neuronal development, neuronal metabolism, neurotransmitter synthesis, and synaptic function. They play a critical role in repair and neurogenesis during CNS and spinal cord injury, and astrocyte dysfunction has also been implicated in many debilitating CNS disorders such as Parkinson’s disease (PD), Alzheimer’s disease, Amyotrophic lateral sclerosis, and Huntington’s disease.
Applied StemCell has two isogenic panels of neuronal lineage cells (neural stem cells, astrocyte precursors & mature cells, dopaminergic neurons) derived from control iPSC lines, ASE-9109 and ASE-9110, that are ideal for comparative evaluation of drug efficacy, toxicity, gene profiling, and construction of complex, functional in vitro neurological models.
Applied StemCell’s iPSC-derived Astrocyte Precursor and Mature Cells are derived from neural stem cells differentiated from fully characterized, normal karyotype human induced pluripotent stem cells (hiPSCs), using proprietary induction protocols that produce high purity astrocytes (>90% GFAP and < 1% Tuj1 cells). The astrocyte precursor cells can be further differentiated into mature astrocytes using the Astrocyte Differentiation Media (ASE-9322MDM) and maintained using our Astrocyte Maintenance Media (ASE-9322MCM).
Advantages of choosing ASC’s iPSC-derived astrocytes:
- >90% GFAP+ cells; <1% Tuj1+ cells with typical astrocyte morphology
- Two astrocyte lineages derived from one male (ASE-9109) and one female (ASE-9110) human iPSC line
- Expression of astrocyte markers confirmed by immunocytochemistry and whole genome profiling
- Suitable for co-culturing with neurons for complex tissue modeling
- Limited proliferative potential and long-term viability
- Xeno-free & Integration-free derivation and culturing
Benefits & Applications:
- Physiologically relevant models for studying neurogenesis and CNS function
- Reliable and consitent source of in vitro models for drug screening & neurotoxicity tests
- Build disease models of neurodegenerative diseases and neuroinflammation
- Co-culture with neurons to improve neuronal viability in cell therapy studies
Also available: Isogenic dopaminergic neurons, and other CNS neurons from the each control iPSC line. Applied StemCell's catalog also provides Astrocyte Precursor Starter Kit and Astrocyte Mature Starter Kit to generate and maintain mature astrocytes from astrocyte precursor cells in your own lab.. The kits include cryopreserved precursors or mature astrocytes, respectively, as well as optimized medium and supplements.
Figure 1. (Left) Astrocyte morphology 17 days post-seeding. (Right) Immunocytochemical characterization of astrocytes derived from NSCs (ASE-9234) shows >90% GFAP+ cells (astrocyte marker) and <1% Tuj1+ cells (neuronal marker).
Figure 2. Whole genome profiling for markers expressed by astrocytes derived from NSCs.
Custom iPSC-derived Astrocyte Differentiation Service We can differentiate astrocytes and other glial cells from your control/ patient iPSC lines or from engineered iPSC lines to suit your project needs, including full characterization of your derived cell lines.
Isogenic neural stem cell derivatives differentiated from a single iPSC line
Applied StemCell's Neural Stem Cells (NSCs) can be differentiated to neuron products including neurons and glial cells (dopaminergic and mixed neurons, astrocytes and oligodendrocytes) using ASC's differentiation and maturation media and kits. NSCs used for further neural lineage cell derivation uniformly express >90% Sox1 and >98% Nestin. These neural stem cell-derived neurons and astrocytes have been used extensively for screening assays in publications.
Co-culturing of neurons and astrocytes to develop complex models
Neurons and astrocytes derived from Applied StemCell's NSCs can be co-cultured for developing complex research models. The cells can be isogenic differentiated cells or cells from different genetic backgrounds.
Figure. Enhanced synpase formation in neuron-astrocyte co-cultures. Co-culture of neurons and astrocytes showed a significant increase in synaptic puncta, as seen by the co-colocalization of neuronal marker (Tuj1) and synapse marker (Synapsin),as compared to neuron only cultures.
Frequently Asked Questions
1. For how many passages can mature astrocytes be cultured?
Mature astrocytes tend to proliferate slowly and we don’t passage the mature astrocytes more than 2 passages after they reach day 35. These will be healthy for at least 4 weeks. In our lab, we have grown healthy and functionally viable astrocytes up to day 80.
2. Do you characterize Astrocyte precursor cells using CD44 or A2B5 staining?
No, we don’t characterize for staining during astrocyte precursor stage. We characterize mature astrocytes using immunohistochemical analysis for GFAP/Tuj1 at day 17 of differentiation.
3. Can astrocyte precursor cells (APCs) be passaged?
Astrocyte precursor cells are intermediate stage cells that are not stable and once thawed, cannot be maintained or passaged at this stage without maturation.
4. Do you have a recommendation for how to determine if the APCs have fully matured?
To determine if the APCs have fully matured, we perform immunohistochemical analysis for GFAP, and also observe if the cells are close to or at their end of their proliferation stage.
5. Is there a separate medium for maintaining astrocyte precursor cells (APCs) in culture?
Astrocyte precursor cells are intermediate stage cells that are not stable and once thawed, cannot be maintained or passaged at this stage without maturation. During the maturation phase, APCs will proliferate and reach confluency when cultured in our proprietary Astrocyte Maturation Medium (ASE-9322AM). At 80-90% confluency, sub-culture the APCs into a new dish until completion of maturation on day 17 of differentiation.
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Efthymiou, A. G., Steiner, J., Pavan, W. J., Wincovitch, S., Larson, D. M., Porter, F. D., ... & Malik, N. (2015). Rescue of an in vitro neuron phenotype identified in Niemann-Pick disease, type C1 induced pluripotent stem cell-derived neurons by modulating the WNT pathway and calcium signaling. Stem cells translational medicine, 4(3), 230-238.
Efthymiou, A., Shaltouki, A., Steiner, J. P., Jha, B., Heman-Ackah, S. M., Swistowski, A., ... & Malik, N. (2014). Functional screening assays with neurons generated from pluripotent stem cell–derived neural stem cells. Journal of biomolecular screening, 19(1), 32-43.
Shaltouki, A., Peng, J., Liu, Q., Rao, M. S., & Zeng, X. (2013). Efficient generation of astrocytes from human pluripotent stem cells in defined conditions. Stem cells, 31(5), 941-952.