Molecular tracking of interactions between progenitor and endothelial cells via Raman and FTIR spectroscopy imaging: a proof of concept of a new analytical strategy for in vitro research

• Published in: Cellular and molecular life sciences : CMLS Vol. 80; no. 11; p. 329
• Main Authors: Augustyniak, Karolina, Pragnaca, Aleksandra, Lesniak, Monika, Halasa, Marta, Borkowska, Agata, Pieta, Ewa, Kwiatek, Wojciech M., Kieda, Claudine, Zdanowski, Robert, Malek, Kamilla
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01.11.2023; Springer Nature B.V; Springer Verlag
• Subjects: Alternations; Animals; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bone marrow; Brain; Cell Biology; Cells, Cultured; Cluster analysis; Cytoplasm; Droplets; Endothelial cells; Endothelial Cells - metabolism; Endothelium; Fourier transforms; Infrared spectroscopy; Life Sciences; Lipid Droplets - metabolism; Lipids; Medical imaging; Mice; Microenvironments; Neuroimaging; Original; Original Article; Osteoprogenitor cells; Progenitor cells; Regression analysis; Spectral signatures; Spectroscopy, Fourier Transform Infrared; Spectrum Analysis, Raman - methods; Multivariate analysis; Mouse brain microvascular endothelial cells (MBrMEC); Mouse aorta gonad mesonephros endothelial cells 11.5 (MAgEC11.5); Intercellular interactions; Raman and FTIR imaging
• ISSN: 1420-682X; 1420-9071
• DOI: 10.1007/s00018-023-04986-3

Circulating endothelial cell progenitors originating from the bone marrow are considered to be a powerful tool in the repair of endothelium damage. Due to their unique properties, endothelial progenitors are now broadly investigated to assess their clinical significance in diseases e.g., associated with brain endothelial dysfunction. However, their distinction in terms of the expression of specific markers remains ambiguous. Additionally, endothelial progenitor cells may change their repertoire of markers depending on the microenvironment of the tissue in which they are currently located. Here, we applied the label-free Raman and FTIR imaging to discriminate mice brain endothelium and endothelial progenitors. Cells cultured separately showed distinctly different spectral signatures extracted from the whole cellular interior as well as the detected intracellular compartments (nucleus, cytoplasm, perinuclear area, and lipid droplets). Then, we used these spectroscopic signals to examine the cells co-cultured for 24 h. Principal cluster analysis showed their grouping with the progenitor cells and segregation from brain endothelium at a level of the entire cell machinery (in FTIR images) which resulted from biochemical alternations in the cytoplasm and lipid droplets (in Raman images). The models included in partial least square regression indicated that lipid droplets are the key element for the classification of endothelial progenitor-brain endothelial cells interactions.