Millifluidic culture improves human midbrain organoid vitality and differentiation
Millifluidic culture improves human midbrain organoid vitality and differentiation
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DOI：
10.1039/c8lc00206a
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发表时间：
2018-10-21
期刊：
影响因子：
6.1
通讯作者：
Schwamborn, Jens C.
中科院分区：
文献类型：
--
作者：
Berger, Emanuel;Magliaro, Chiara;Schwamborn, Jens C.
人类中脑特异性类器官（hMOs）是研究帕金森病（PD）发病机制的实验体外模型。在hMOs中，神经上皮干细胞（NESCs）产生功能性中脑多巴胺能（mDA）神经元，这些神经元在PD期间选择性变性。hMO模型的一个局限性是向密集的核心区提供的氧气和营养物质不足，这最终导致一个死核。为了减少这种现象，我们采用了一种微流体培养系统，通过连续的层流来保证介质的供应。我们开发了一个氧气运输和消耗的计算模型，以预测hmo内的氧气水平。该模型预测，在微流体条件下，hMO核心区域的氧含量较高。与计算模型一致的是，在生物反应器系统中培养的hMOs与在常规摇晃条件下培养的hMOs相比，死核明显更小。将类器官中坏死的核心区域与从模型中获得的区域进行比较，可以估计确保细胞活力所需的临界氧浓度。除了减少死核大小外，中流处理的hMOs从NESCs向mDA神经元的分化效率也有所提高。分化的增加涉及代谢成熟过程，在微流体培养中进一步发展。总的来说，在先进PD建模的背景下，提高hMO质量的生物反应器条件值得考虑。
Human midbrain-specific organoids (hMOs) serve as an experimental in vitro model for studying the pathogenesis of Parkinson's disease (PD). In hMOs, neuroepithelial stem cells (NESCs) give rise to functional midbrain dopaminergic (mDA) neurons that are selectively degenerating during PD. A limitation of the hMO model is an under-supply of oxygen and nutrients to the densely packed core region, which leads eventually to a dead core. To reduce this phenomenon, we applied a millifluidic culture system that ensures media supply by continuous laminar flow. We developed a computational model of oxygen transport and consumption in order to predict oxygen levels within the hMOs. The modelling predicts higher oxygen levels in the hMO core region under millifluidic conditions. In agreement with the computational model, a significantly smaller dead core was observed in hMOs cultured in a bioreactor system compared to those ones kept under conventional shaking conditions. Comparing the necrotic core regions in the organoids with those obtained from the model allowed an estimation of the critical oxygen concentration necessary for ensuring cell vitality. Besides the reduced dead core size, the differentiation efficiency from NESCs to mDA neurons was elevated in hMOs exposed to medium flow. Increased differentiation involved a metabolic maturation process that was further developed in the millifluidic culture. Overall, bioreactor conditions that improve hMO quality are worth considering in the context of advanced PD modelling.