Table 1.

Comparison of typical neural culture systems.

Culture System	Application	Pros	Cons
Organotypic slices	- Complex neural circuit modeling - Tumor invasion analysis	Retain in vivo organization	- Limited culture life - Genetic manipulation difficult
Primary neurospheres	- Neural stem and progenitor cell expansion - Tumor cell expansion	- Isolation of mature (active) cell types and pure cell populations possible - Co-culture of distinct cell types possible - Low variability between experiments - Ease of expansion for high cell yield	- Limited passage number - Technically difficult to passage - Need sufficient starting cell source - Difficult to establish from adult tissues - Lacks in vivo organization and diverse cell types
Primary cell adherent culture	- Separating cell autonomous from non-autonomous effects	- Isolation of mature (active) cell types and pure populations of some cell types - Co-culture of distinct cell types possible - Low variability between experiments	- Isolating pure cell types difficult and time consuming - Difficult to establish from adult tissues - Genetic manipulation difficult outside of proliferating NPCs - Limited culture life-span for mature cell types - Lacks in vivo organization and diverse cell types
hPSC-derived adherent culture	- Separating cell autonomous from non-autonomous effects - Human genetic studies (background variability and single gene mutations) - Neurogenesis and gliogenesis	- Accessible human relevant cells - Pure culture and co-cultures possible - Can generate specific cell subtypes - Easy to produce multiple cell types from one initial source - Early proliferating cell types (PSCs or NPCs) are expandable and bankable - Genetic manipulation protocols well established	- Cells produced are typically fetal-like - Lacks in vivo organization - Long differentiation protocols and maturation times - Mastery of hPSC culture a prerequisite - High cell line to cell line variability
hPSC-derived spheroids and organoids	Tumor cell expansion, Modeling brain development and neurodevelopmental disorders	- High efficiency of neural differentiation - Observe interactions between multiple cell types - In vivo-like organization - Spontaneous (unpatterned) or directed (patterned) differentiation possible	- Similar to adherent culture - Lacking some critical non-CNS cell types (e.g., microglia and vasculature), though these can be added by co-culture