High cell density three-dimensional neural co-cultures require continuous medium perfusion for survival.
High cell density three-dimensional neural co-cultures require continuous medium perfusion for survival.
复制标题
高细胞密度三维神经共培养物需要连续的培养基灌注才能存活。
DOI：
10.1109/iembs.2006.260639
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
LaPlaca,MichelleC
中科院分区：
文献类型：
--
作者：
Cullen,DKacy;Vukasinovic,Jelena;Glezer,Ari;LaPlaca,MichelleC
与传统的二维 (2-D) 培养模型相比，神经细胞培养的三维 (3-D) 模型可以为研究人员提供更具生理相关性的环境来研究神经生物学现象。然而，在厚（> 500mum）3-D 培养物的开发中，扩散限制了质量运输，因此需要使用远低于中枢神经系统（CNS）中的细胞密度。本研究的目的是评估连续培养基灌注对细胞密度接近脑组织中的厚 3-D 神经元-星形胶质细胞共培养物存活的影响。在这些研究中使用的细胞密度和厚度分别为 104 个细胞/mm3 和 500-750mum 时，非灌注培养物表现出广泛的细胞/基质降解和细胞死亡。然而，与非灌注共培养物相比，以相对较高的速率（2.5-11.0μL/分钟，对应于每天6-27次培养基交换）灌注的共培养物表现出降解减少和活力增强。此外，评估的最高灌注速率为 11.0μL/min，可实现 >90% 的细胞活力和培养厚度的维持。下一代 3D 神经培养物的细胞类型和密度更接近中枢神经系统，可以提供增强的模型保真度，并且在细胞生长、相互作用以及对化学或机械扰动的反应的机制研究中有价值
Three-dimensional (3-D) models of neural cell culture may provide researchers with a more physiologically-relevant setting to study neurobiological phenomena than traditional two-dimensional (2-D) culture models. However, in the development of thick (>500mum) 3-D cultures, diffusion limited mass transport necessitated the use of cell densities much lower than those found in the central nervous system (CNS). The goal of this study was to evaluate the effects of continuous medium perfusion on the survival of thick, 3-D neuronal-astrocytic co-cultures at cell densities closer to those found in brain tissue. At the cell density and thickness used for these studies, 104cells/mm3and 500-750mum, respectively, non-perfused cultures exhibited widespread cellular/matrix degradation and cell death. However, co-cultures perfused at relatively high rates (2.5-11.0muL/min, corresponding to 6-27 medium exchanges/day) demonstrated decreased degradation and enhanced viability compared to non-perfused co-cultures. Furthermore, the highest perfusion rate evaluated, 11.0muL/min, resulted in >90% cell viability and maintenance of culture thickness. Next generation 3-D neural cultures, with cell types and densities better approximating the CNS, may provide enhanced model fidelity and be valuable in the mechanistic study of cell growth, interactions, and the responses to chemical or mechanical perturbations