Differentiation of Multiple iPSC Lines to Different Cell Types to Enable Cohort Analysis in Neurodegeneration: Challenges and Learnings

The poster Differentiation of Multiple iPSC Lines to Different Cell Types to Enable Cohort Analysis in Neurodegeneration: Challenges and Learnings presents Axol Bioscience’s approach to building scalable, human-relevant in vitro models using multiple patient-derived iPSC lines.

Key highlights include:

• Cohort-Level Differentiation: Eight axoLines™ iPSC lines (2 healthy, 6 Alzheimer’s disease) with diverse genotypes (ApoE3/E4, ApoE4/E4, PSEN1 mutations) were differentiated into astrocytes, cortical excitatory neurons, and microglia.
• Standardized QC:
  • iPSCs: Sterility, morphology, pluripotency markers (SSEA4, OCT3/4, TRA-1-60), karyotyping, Sendai clearance
  • Neurons: BRN2, CUX1, TUJ1, MAP2; absence of GFAP and OLIG2
  • Astrocytes: GFAP, Nestin, EAAT2, S100β, ALDH1, AQP4, TUJ1
  • Microglia: TMEM119, Iba1, P2RY12, CX3CR1
• Optimization Protocols:
  • Astrocyte differentiation adjusted for passaging ratios and maturation timing
  • Cryopreservation of progenitors enables repeatable workflows
  • Microglia yield scaled by flask size and density
• Marker Validation: Immunocytochemistry confirmed cell-type-specific marker expression across all lines.
• Future Directions: Functional assays including phagocytosis, cytokine release, and spontaneous firing in triculture systems will further characterize phenotypic differences.

This cohort-based strategy supports the development of complex neurodegenerative disease models for compound screening and personalized medicine.

Featured Axol Products:

• axoCells™ Astrocytes
• axoCells™ Cortical Excitatory Neurons
• axoCells™ Microglia