Table 2.

Strategies to overcome the “necrotic core” in brain cultures, with their pros and cons.

	Benefits	Limitations	References
Shaking cultures and micro-fluidic devices	Reduced cell death Shaking cultures easy and cheap Enabling continuous medium flow	Do not prevent the formation of the necrotic core	Berger et al. (2018)
Sliced organoids	Recapitulates mature cortical cells subtypes Expression of ALS pathogenic markers in both glia and neurons	Repeated slicing may impair axonal outgrowth 3D partially lost	Szebényi et al. (2021)
Air-liquid interface	Reduced cell death Preserved axonal outgrowth Functional neural circuits present	Late embryonic structures preserved Cortical layers not identifiable 3D partially lost	Giandomenico et al. (2019)
Organoids-on-chip	Functional oxygenation	Expensive method	Salmon et al. (2022)
Direct reprogramming	Improved neuronal functionality Formation of BBB-like structures	Lack of reproducibility	Cakir et al. (2019)
Vascular organoids	Formation of endothelial tubes with smooth muscle cells, pericytes and basement membrane ·Establishment of a network between vessels and neural cells	Lack of BBB specific permeability Lack of a systemic flow	Wimmer et al. (2019)
Hyperoxygenation	Reduced cell death	Physiological condition not represented	Watanabe et al. (2017)
Transplantation in animal models	Reduced cell death Host vascularization	Physiological condition not represented	Popatansov (2018)