The field of tissue engineering has not yet provided knowledge on which a consensus for the complex physiological relevance (CPR) of neuronal cultures could be established. The CPR of 3D neuronal cultures can have a profound impact on the drug discovery process through the validation of in vitro models for the study of neuropsychiatric and degenerative diseases, as well as screening for neurotoxicity during drug development. Herein, we assemble evidence in support of the potential of [Ca²⁺]_i oscillation frequency as a CPR outcome that can demonstrate the in vivo-like behavior of 3D cultures and differentiate them from 2D monolayers. We demonstrate that [Ca²⁺]_i oscillation frequencies in 2D cultures are significantly higher than those found in 3D cultures, and provide a possible molecular explanation.

组织工程领域尚未提供知识，可以就神经元培养物的复杂生理相关性（CPR）达成共识。通过验证用于研究神经精神病和退行性疾病的体外模型以及筛选药物开发过程中的神经毒性，3D神经元文化的CPR可以对药物发现过程产生深远影响。本文中，我们收集了支持[Ca ²⁺ ] _i振荡频率作为CPR结果的证据，该结果可以证明3D培养物的体内样行为，并将它们与2D单层细胞区分开。我们证明了[Ca ²⁺ ] _i 2D文化中的振荡频率明显高于3D文化中的振荡频率，并提供了可能的分子解释。