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Updated: Jul 7, 2026
Chemogenetic Regulation in Reprogrammed Stem Cell-derived Precursor Cells in Treating Neurodegenerative Diseases
Published on: May 2, 2025
Artificial cerebrospinal fluid preserves neuronal viability and attenuates apoptosis and oxidative stress in HT22
Lingyi Chi1, Liangwen Zhang2, Zhiyong Ma3
1Department of Neurosurgery, Qilu Hospital, Shandong University, Jinan, Shandong, China.
Introduction:
Artificial cerebrospinal fluid (ACSF) is critical for maintaining neuronal function during surgical procedures, yet its specific neuroprotective advantages over commonly used irrigation solutions like physiological saline and phosphate-buffered saline (PBS) remain unclear.
Methods:
This study compared the neuronal preservation properties of ACSF, saline, and PBS using HT22 mouse hippocampal neurons, with DMEM as a control. Cell viability and apoptosis were assessed using CCK-8 assays and Annexin V-FITC/PI flow cytometry. Mitochondrial morphology and function were evaluated by transmission electron microscopy and mitochondrial membrane potential (ΔΨm) measurements. Intracellular reactive oxygen species (ROS) and calcium levels were determined using DCFH-DA and Fura-2 AM staining. Key signaling pathways were analyzed via Western blotting.
Results:
Results showed that ACSF significantly preserved cell viability and reduced apoptosis compared to saline and PBS. This effect was mediated by preserving mitochondrial integrity and ΔΨm, reducing the Bax/Bcl-2 ratio, suppressing ROS accumulation, and maintaining calcium homeostasis. Mechanistically, ACSF specifically inhibited the p38 MAPK/MK2 and JNK stress pathways without affecting NF-κB.
Discussion:
We conclude that ACSF offers superior preservation of neuronal viability compared with saline-based solutions by preserving mitochondrial function, reducing oxidative stress, and inhibiting key apoptotic pathways, supporting its preferential use in neurosurgical applications, pending in vivo validation.
