Islam Rezwanul, Choudhary Hadi Hasan, Mehta Hritik, Zhang Feng, Jovin Tudor G, Hanafy Khalid A
Department of Biomedical Sciences, Cooper Medical School at Rowan University, Camden, NJ, USA.
Cooper Neurological Institute, Cooper University Health Care, Camden, NJ, USA.
Transl Stroke Res. 2025 Jun;16(3):655-671. doi: 10.1007/s12975-024-01243-y. Epub 2024 Apr 1.
Subarachnoid hemorrhage (SAH) accounts for 5% of stroke, with women having a decreased inflammatory response compared to men; however, this mechanism has yet to be identified. One hurdle in SAH research is the lack of human brain models. Studies in murine models are helpful, but human models should be used in conjunction for improved translatability. These observations lead us to develop a 3D system to study the sex-specific microglial and neuroglial function in a novel in vitro human SAH model and compare it to our validated in vivo SAH model. Our lab has developed a 3D, membrane-based in vitro cell culture system with human astrocytes, microglia, and neurons from both sexes. The 3D cultures were incubated with male and female cerebrospinal fluid from SAH patients in the Neuro-ICU. Furthermore, microglial morphology, erythrophagocytosis, microglial inflammatory cytokine production, and neuronal apoptosis were studied and compared with our murine SAH models. The human 3D system demonstrated intercellular interactions and proportions of the three cell types similar to the adult human brain. In vitro and in vivo models of SAH showed concordance in male microglia being more inflammatory than females via morphology and flow cytometry. On the contrary, both in vitro and in vivo models revealed that female microglia were more phagocytic and less prone to damaging neurons than males. One possible explanation for the increased phagocytic ability of female microglia was the increased expression of CD206 and MerTK. Our in vitro, human, 3D cell culture SAH model showed similar results to our in vivo murine SAH model with respect to microglial morphology, inflammation, and phagocytosis when comparing the sexes. A human 3D brain model of SAH may be a useful adjunct to murine models to improve translation to SAH patients.
蛛网膜下腔出血（SAH）占中风病例的5%，与男性相比，女性的炎症反应有所降低；然而，这一机制尚未明确。SAH研究中的一个障碍是缺乏人脑模型。小鼠模型研究很有帮助，但应结合使用人体模型以提高可转化性。这些观察结果促使我们开发一种三维系统，在一种新型的体外人体SAH模型中研究性别特异性小胶质细胞和神经胶质细胞功能，并将其与我们经过验证的体内SAH模型进行比较。我们实验室开发了一种基于膜的三维体外细胞培养系统，其中包含来自男女两性的人类星形胶质细胞、小胶质细胞和神经元。将三维培养物与神经重症监护病房中SAH患者的男性和女性脑脊液一起孵育。此外，还研究了小胶质细胞形态、红细胞吞噬作用、小胶质细胞炎性细胞因子产生和神经元凋亡，并与我们的小鼠SAH模型进行了比较。人体三维系统显示出细胞间相互作用以及三种细胞类型的比例与成人大脑相似。SAH的体外和体内模型通过形态学和流式细胞术显示出一致的结果，即雄性小胶质细胞比雌性更具炎症性。相反，体外和体内模型均显示，雌性小胶质细胞比雄性更具吞噬作用，且对神经元的损伤倾向更小。雌性小胶质细胞吞噬能力增强的一个可能解释是CD206和MerTK表达增加。在比较性别时，我们的体外人体三维细胞培养SAH模型在小胶质细胞形态、炎症和吞噬作用方面与我们的体内小鼠SAH模型显示出相似的结果。SAH的人体三维脑模型可能是小鼠模型的有用辅助工具，有助于更好地转化应用于SAH患者。
