PhD Scientific Days 2025

Poster Session III. - R: Neurosciences

Changes in Neuronal Network and Glial Activation in Human Organotypic Brain Slice Cultures

Name of the presenter

Stelcz Rebeka

Institute/workplace of the presenter

HUN-REN Research Centre for Natural Sciences

Authors

Rebeka Stelcz^1,2, Nour Essam Fawzy Ahmed Aly^1,3, Réka Bod^1,2, Eszter Juhász¹, Kinga Tóth¹, Estilla Zsófia Tóth⁴, István Ulbert^1,3,5, Lucia Wittner^1,2,5

1: HUN-REN Research Centre for Natural Sciences
2: Semmelweis University, János Szentágothai Neurosciences Division
3: PPCU, Roska Tamás Doctoral School of Sciences and Technology
4: Hun-Ren Research Centre for Natural Sciences
5: Semmelweis University, Department of Neurosurgery

Text of the abstract

Introduction: Several animal and computational models help study the central nervous system's properties, advancing our understanding of neuronal behaviour, neurodegenerative diseases, and potential treatments. Despite their value, translating findings from animal models to humans is limited by the human brain's unique cellular, molecular, and network features. Human organotypic brain tissue cultures offer an excellent opportunity to explore the characteristics of the human brain on the cellular and molecular level and assess novel pharmaceutical compounds in a more realistic environment.
Aims: In this study, we explored neuronal degeneration and loss, as well as the degree of glial activity along the culture period.
Method: Postoperative brain tissues were sliced and maintained in six-well plates under sterile conditions. Two culture media (artificial culturing medium, human cerebrospinal fluid) and distinct antibiotics, antimycotics (penicillin, streptomycin, amphotericin) for infection prevention were tested. Morphological modifications of neurons were investigated using immunostaining against the neuronal marker NeuN and the dendrite marker MAP2. Changes in astrocyte and microglial activation patterns were detected with anti-GFAP and anti-IBA1 staining, respectively. These methods enabled us to assess the extent to which the cytoarchitecture of the neocortical tissue culture is preserved in comparison to its acute state regarding the neuronal cell density, morphology and glial activation.
Results: We found that while organotypic cultures maintain overall tissue architecture, considerable changes occur over time. Neuronal density was decreasing mostly in the early culturing period, while astroglial and microglial activation was seen at the longer term during the culturing period.
Conclusion: According to the results, although human organotypic cultures preserve their structure, they show remarkable reorganization with time, highlighting the importance of continous anatomical examinations of the cultures to better understand the morphological modifications and their relation to the physiology of these slice cultures.

Funding:
National Research, Development and Innovation Office K137886 and Bolyai Fellowship
Hungarian Brain Research Program NAP2022-I-8/2022,
PharmaLab RRF-2.3.1-21-2022-00015
NKFIH Advanced 150799, OTKA K137886