Ene Justice, Muok Laureana, Gonzalez Vanessa, Sanchez Nicolas, Nathani Aakash, Syed Falak, Liu Zixiang Leonardo, Singh Mandip, Driscoll Tristan, Li Yan
Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL 32310, USA.
College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL 32310, USA.
Biomedicines. 2025 Apr 28;13(5):1069. doi: 10.3390/biomedicines13051069.
Choroid plexus is a complex structure in the human brain that is responsible for the secretion of extracellular vesicles (EVs) in cerebrospinal fluid. Few studies to date have generated choroid plexus (ChP) organoids differentiated from human induced pluripotent stem cells (hiPSCs) and analyzed their secreted EVs. The scalable Vertical-Wheel bioreactors (VWBRs) provide low shear stress and a controlled environment. This study utilized VWBRs for the differentiation of hiPSCs into ChP organoids and generation of the secreted EVs compared to a static culture. Additionally, this study loaded curcumin into ChP organoid-derived EVs, performed EV lyophilization, and determined the ability of the re-hydrated EVs to alleviate neuro-inflammation. The results demonstrated that the VWBR culture exhibited more aerobic metabolism and active glucose and glutamine consumption than the static control. Consequently, the ChP markers and Endosomal Sorting Complexes Required for Transport-dependent and -independent EV biogenesis genes were significantly upregulated (2-3-fold) in the VWBR, producing four-fold-higher EVs per mL media than the static control. The EVs retained similar size and zeta potential after lyophilization and re-hydration. The cells exposed to amyloid beta 42 oligomers and treated with the curcumin-loaded re-hydrated EVs showed high viability and the reduced inflammatory response determined by TNF-α and IL-6 expression. This study demonstrates a scalable bioreactor system to promote ChP organoid differentiation and generation of EV-based cell-free therapeutics to treat neural inflammation in various neurological disorders.
脉络丛是人类大脑中的一种复杂结构，负责脑脊液中细胞外囊泡（EVs）的分泌。迄今为止，很少有研究从人诱导多能干细胞（hiPSCs）中分化出脉络丛（ChP）类器官并分析其分泌的EVs。可扩展的垂直轮式生物反应器（VWBRs）提供低剪切应力和可控环境。与静态培养相比，本研究利用VWBRs将hiPSCs分化为ChP类器官并生成分泌的EVs。此外，本研究将姜黄素加载到ChP类器官衍生的EVs中，进行EV冻干，并确定复水后的EVs减轻神经炎症的能力。结果表明，与静态对照相比，VWBR培养表现出更多的有氧代谢以及活跃的葡萄糖和谷氨酰胺消耗。因此，VWBR中ChP标志物以及运输依赖性和非依赖性EV生物发生基因所需的内体分选复合物显著上调（2至3倍），每毫升培养基产生的EVs比静态对照高四倍。冻干和复水后，EVs保持相似的大小和zeta电位。暴露于淀粉样β42寡聚体并用负载姜黄素的复水EVs处理的细胞显示出高活力，并且通过TNF-α和IL-6表达确定炎症反应降低。本研究展示了一种可扩展的生物反应器系统，以促进ChP类器官分化并生成基于EV的无细胞疗法，用于治疗各种神经系统疾病中的神经炎症。
