✍🏼 Boglarka Csik, Ádám Nyúl-Tóth, Rafal Gulej, Roland Patai, Tamas Kiss, Jordan Delfavero, Raghavendra Y. Nagaraja, Priya Balasubramanian, Santny Shanmugarama, Anna Ungvari, Siva Sai Chandragiri, Kiana Vali Kordestan, Mark Nagykaldi, Peter Mukli, Andriy Yabluchanskiy, Sharon Negri, Stefano Tarantini, Shannon Conley, Tae Gyu Oh, Zoltan Ungvari, Anna Csiszar

🏠 Vascular Cognitive Impairment, Neurodegeneration, and Healthy Brain Aging Program, Department of Neurosurgery, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA

📑 Aging Cell (2025)

Abstract
With advancing age, neurovascular dysfunction manifests as impaired neurovascular coupling (NVC), microvascular rarefaction, and blood–brain barrier (BBB) disruption, contributing to vascular cognitive impairment (VCI). Our previous research established a causal link between vascular senescence induced cerebromicrovascular dysfunction and cognitive decline in accelerated aging models. The present study examines whether chronological aging promotes endothelial senescence, adversely affecting neurovascular health, and whether senolytic therapies can enhance neurovascular function and cognitive performance in aged mice. We used transgenic p16-3MR mice to identify and eliminate senescent cells and employed genetic (ganciclovir) and pharmacological (ABT263/Navitoclax) senolytic approaches. Evaluations included spatial memory performance, NVC responses, cortical microvascular density, BBB permeability, and detection of senescent endothelial cells via flow cytometry. Brain endothelial cells exhibited heightened sensitivity to aging-induced senescence, undergoing senescence at a greater rate and earlier than other brain cell types, particularly during middle age. This microvascular endothelial cell senescence was associated with NVC dysfunction, microvascular rarefaction, BBB disruption, and deteriorating cognitive performance. On the other hand, senolytic treatments in aged mice improved NVC responses, BBB integrity, microvascular density, and learning capabilities. Notably, these findings suggest that the most effective time window for senolytic treatment is in middle-aged mice, where early intervention could better prevent neurovascular dysfunction and mitigate age-related cognitive impairment.

How the WOLF was used in this study
In the Aging Cell article (DOI: 10.1111/acel.70048), the authors used the WOLF Cell Sorter (NanoCellect) as part of their preparation of brain-derived single-cell suspensions for downstream molecular analysis. After tissue digestion and debris removal, they stained cells to identify live cells and then applied the WOLF Cell Sorter to perform low-pressure fluorescent activated cell sorting (FACS), efficiently removing dead cells and cellular debris to enrich the sample for viable, high-quality single cells. This enriched cell population was critical for preserving cell integrity ahead of transcriptomic sequencing and related analyses, ensuring that the resulting data reflect healthy, intact cells suitable for transcriptome profiling.