AMSBIO-Biologically relevant in vitro models for neuroscience research

AMSBIO-Biologically relevant in vitro models for neuroscience research

2018.02.13

 IPSC Derived Neural Cells

amsbio now offers a new range of Neural Stem Cells (NSCs) and neural lineage progenitors (astrocytes, cortical neurons and dopaminergic neurons) are derived from control human iPSCs using non-integrating episomal-based differentiation methods. Also, available are optimized media and kits for easy differentiation to high quality neural lineage cells.

Neural Stem Cells

amsbio offer ready-to-use Neural Stem Cells (NSCs) and neural lineage progenitors (astrocytes, cortical neurons and dopaminergic neurons) are derived from control human iPSCs using non-integrating episomal-based differentiation methods. Also, available are optimized media and kits for easy differentiation to high quality neural lineage cells.

Benefits:

  • Different NSC lines, each derived from a single control iPSC line or engineered iPSC line: male (cord blood), female (cord blood), male (fibroblasts) and more
  • High purity (>90%) cells expressing neural stem cell markers: SOX1, PAX6 and Nestin
  • Fully characterized by immunocytochemistry and whole genome profiling
  • Provides a consistent and reliable source of differentiated mature, functional neurons and glial cells
  • Physiologically relevant cell line models

 

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Neural Stem Cells

Fig 1. Neural stem cells (ASE-9303) differentiated from a well-characterized control fibroblast-derived iPSC line (ASE-9203) using these proprietary neural induction protocol and NSC culture media. These differentiated NSCs retain neural stem cell phenotype even after passaging and freeze-thaw under feeder-free conditions as evidenced by staining for NSC markers PAX6 (>90%) and SOX1 (>90%) and nuclear DNA (DAPI). Note: White arrows indicate distinctive neuronal rosettes.

 

Astrocytes

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.

Benefits:

  • 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



 

amsbio supplies 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.

These 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).

 

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Astrocytes

Fig 2. 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.

 

Dopaminergic Neurons

For several years, neuroscience research using cell line and animal models of degenerative neurological disorders such as Parkinson’s disease (PD) and Alzheimer’s has been plagued with the lack of translation of results to the human scenario, whether for basic understanding of disease pathology or for screening of targeted drug therapies. The development of the induced pluripotent stem cell (iPSC) technology and its further differentiation into various post-mitotic somatic cell lineages, especially neuronal lineage cells and dopaminergic neurons, has overcome a major bottleneck by providing biologically relevant in vitro models to screen drugs for neuroprotection and movement-disorder therapy.

Benefits:

  • Yields high purity, functional dopamine neurons after maturation
  • Mature dopamine neurons can be maintained long-term in culture (up to 3 weeks)
  • Differentiated from NSCs using footprint-free, integration-free protocols
  • Two dopaminergic lineages (ASE-9323, ASE-9323F): derived from one male donor (ASE-9109) and one female donor (ASE-9110)
  • Also, available: isogenic panels of neural stem cells, other differentiated neurons, astrocytes precursors and mature cells from same donor iPSCs
  • Proven as an ideal model for disease modeling, neuroprotection and neurotoxicity screening assays



 

amsbio supplies Human Dopaminergic neurons cryo-preserved, pre-differentiated dopaminergic neuron precursors derived from a footprint-free, karyotype normal human iPSC line. These precursors yield high purity mature dopaminergic neurons using these optimized dopaminergic maturation medium and supplements (ASE-9323DM). Mature dopaminergic neurons can be obtained within 12 days after thaw, and the matured dopaminergic neurons can be maintained in culture for up to 3 weeks without losing functionality. These mature and functional dopaminergic neurons contain >80% neurons, >30% TH+ dopaminergic neurons and < 15% GFAP+ astrocytes.

 

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Dopaminergic Neurons

Fig 3. Immunocytochemical characterization of mature dopaminergic neurons (ASE-9323) differentiated from iPSC-derived neural stem cells (parent iPSC line: ASE-9109). Top left: Bright field image indicates healthy dopaminergic cell morphology at 12 days post-seeding. Other panels: Immunostaining on day 12 post-seeding shows that >90% of total cells expressed Tuj-1 marker (green) and >30% expressed TH marker (red). A count of the number of nuclei (DAPI; blue) was used to calculate the total number of cells.



 

Mixed Population Neuronal Precursors

amsbio cryo-preserved, pre-differentiated mixed population neuronal precursors derived from a footprint-free, karyotype normal human iPSC line. They yield high purity, functional neurons within 8 days when using these well-optimized maturation medium and supplements. The derived-neurons have also been tested for neuronal activity using functional and screening assays. The neurons can be seeded on various culture vessel formats including 96-well plates on either glass or plastic surfaces and cultured as adherent cells. Shortly after seeding, the cells proliferate slightly for up to 3 days and show extensive neurite outgrowth and proper neuronal morphology. In general, on Day 8 post-seeding, the cell population will contain >90% neurons and < 5% Glial Fibrillary Acidic Protein (GFAP) positive astrocytes.

Benefits:

  • High purity neurons: >90% Tuj1+ neurons and < 5% GFAP+ cells
  • High cell viability of cyropreserved neurons (>80%)
  • Available as isogenic lineages of mixed neurons and associated neuronal derivatives from two different control iPSC lines (male ASE-9109 and female ASE-9110)
  • Functionally viable neurons capable of neuronal excitability and synapse formation
  • Long-term viability in cell culture and consistency in results (>30 days): affords greater flexibility in designing high throughput drug screening and testing

 

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Neuronal Precursors

Fig 4. Immunocytochemical characterization of neurons derived from male cord blood control iPSC line (ASE-9109). Immunocytochemical staining of neurons differentiated from iPSC indicate > 90% of the cells stain positive for neuronal markers Tuj1 (green) and GABA (red); < 5% are GFAP+ cells. indicating high purity of neuronal cell population.

 

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