(1129-B) Differentiation of multiple iPSC lines to different cell types to enable cohort analysis in neurodegeneration: challenges and learnings

Abstract: Differentiation of induced pluripotent stem cells (iPSCs) describes the direction of reprogrammed stem cells, through a specific lineage, into desired endpoint cells (for instance neurons or neuroinflammatory cells). Differentiation of a single stem cell to a single endpoint cell is, for many diseases with complex underlying genetics, not at a sufficient scale to capture the diversity of the underlying condition. Rather, building a cohort-level model, where multiple donor stem cells are derived into multiple end cell types may provide the power to unravel the complexities of conditions such as Alzheimer’s Disease.

We have previously described how iPSC-derived neurons and neuroinflammatory cells can be generated and used together in complex co-culture models for compound screening and drug development (1).

Here we describe the development of differentiation procedures for multiple lines at the same time. We worked on 8 iPSC lines: 2 control lines and 6 from individuals with Alzheimer’s Disease, with mutations in presenilin 1 or with either homozygous ApoE4 or heterozygous ApoE4/ApoE3 genotype.

We differentiated each iPSC into astrocytes, cortical excitatory neurons and microglia and recorded expression of relevant markers (both pluripotency markers of iPSCs and endpoint cell markers for each cell type) highlighting our quality control processes. Standardization of optimization processes is detailed, to drive a platform for robust generation of cells to generate models of neurodegeneration.


1 Steven D. Broadbent, Sian Humphrey, Signe Springe, Sung Min Yang, Ayan Ghoshal, Nina Dedic & Ashley Barnes. Validation of a cortical tri-culture axoModel for in vitro compound screening a blinded compound study