Decoding Cell–Cell Interactions in Neurovascular Unit Co-Cultures Through Molecular Vibrational Spectroscopy and Biology
- 1Faculty of Chemistry, Department of Chemical Physics, Jagiellonian University in Krakow, Gronostajowa 2, 30-387 Krakow, Poland
- 2Doctoral School of Exact and Natural Sciences, Jagiellonian University in Krakow, Prof. S. Lojasiewicza 11, 30-348 Krakow, Poland
- 3Laboratory of Molecular Oncology and Innovative Therapies, Military Institute of Medicine-National Research Institute, Szaserow 128, 04-141 Warsaw, Poland
Abstract
The blood–brain barrier (BBB) is a coordinated multicellular interface in which intercellular communication shapes responses to injury and metabolic stress. However, the contributions of individual cell types to collective stress adaptation remain poorly understood. Here, we examined molecular and metabolic responses of 2D neurovascular unit (NVU) co-culture models comprising endothelial cells, astrocytes, and pericytes exposed to hypoxia or sublethal HgCl2 toxicity. An integrated platform combining gene and protein expression profiling, label-free vibrational spectroscopic imaging, and machine-learning-based analysis was used to assess stress-induced changes in cell-cycle regulation and metabolism across mono- and co-culture configurations. Hypoxia broadly suppressed G1/S and G2/M regulators, inducing cell-cycle arrest with limited cell death, consistent with an adaptive response. HgCl2 caused heterogeneous, model-dependent changes in viability and proliferative capacity. Spectroscopic phenotyping revealed that the triple co-culture exhibited spectral features resembling those of pericyte monocultures, although the contributions of cell–cell interactions and differences in culture conditions could not be distinguished. Fluorescence imaging further revealed mitochondria-associated signals at cell–cell contact sites, suggesting a possible association of mitochondrial material with regions of close cellular interaction. The presence of pericytes was associated with a more stress-resistant cellular phenotype in the investigated co-culture configurations. The developed spectroscopic–molecular platform provides a label-free framework for characterizing cellular communication and stress responses in multicellular NVU co-culture models.
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BibTeX
@article{pragnaca2026decoding,
title = {{Decoding Cell–Cell Interactions in Neurovascular Unit Co-Cultures Through Molecular Vibrational Spectroscopy and Biology}},
author = {Aleksandra Pragnąca and Agata Borkowska and Anna Antolak and Monika Leśniak and Karolina Augustyniak and Iga Czechowska and Kamilla Malek and Robert Zdanowski},
journal = {Cells},
year = {2026},
month = oct,
doi = {10.3390/cells15201873},
url = {https://www.mdpi.com/2073-4409/15/20/1873},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Decoding Cell–Cell Interactions in Neurovascular Unit Co-Cultures Through Molecular Vibrational Spectroscopy and Biology
AU - Aleksandra Pragnąca
AU - Agata Borkowska
AU - Anna Antolak
AU - Monika Leśniak
AU - Karolina Augustyniak
AU - Iga Czechowska
AU - Kamilla Malek
AU - Robert Zdanowski
PY - 2026
DA - 2026/10/15
JO - Cells
DO - 10.3390/cells15201873
UR - https://www.mdpi.com/2073-4409/15/20/1873
AB - The blood–brain barrier (BBB) is a coordinated multicellular interface in which intercellular communication shapes responses to injury and metabolic stress. However, the contributions of individual cell types to collective stress adaptation remain poorly understood. Here, we examined molecular and metabolic responses of 2D neurovascular unit (NVU) co-culture models comprising endothelial cells, astrocytes, and pericytes exposed to hypoxia or sublethal HgCl2 toxicity. An integrated platform combining gene and protein expression profiling, label-free vibrational spectroscopic imaging, and machine-learning-based analysis was used to assess stress-induced changes in cell-cycle regulation and metabolism across mono- and co-culture configurations. Hypoxia broadly suppressed G1/S and G2/M regulators, inducing cell-cycle arrest with limited cell death, consistent with an adaptive response. HgCl2 caused heterogeneous, model-dependent changes in viability and proliferative capacity. Spectroscopic phenotyping revealed that the triple co-culture exhibited spectral features resembling those of pericyte monocultures, although the contributions of cell–cell interactions and differences in culture conditions could not be distinguished. Fluorescence imaging further revealed mitochondria-associated signals at cell–cell contact sites, suggesting a possible association of mitochondrial material with regions of close cellular interaction. The presence of pericytes was associated with a more stress-resistant cellular phenotype in the investigated co-culture configurations. The developed spectroscopic–molecular platform provides a label-free framework for characterizing cellular communication and stress responses in multicellular NVU co-culture models.
SN - 2073-4409
ER -