Details

Autor(en) / Beteiligte

• Radwan, Basseem
• Rocchetti, Stefano
• Matuszyk, Ewelina
• Sternak, Magdalena
• Stodulski, Maciej
• Pawlowski, Robert
• Mlynarski, Jacek
• Brzozowski, Krzysztof
• Chlopicki, Stefan
• Baranska, Malgorzata

Titel

EdU sensing: The Raman way of following endothelial cell proliferation in vitro and ex vivo

Ist Teil von

• Biosensors & bioelectronics, 2022-11, Vol.216, p.114624-114624, Article 114624

Ort / Verlag

Elsevier B.V

Erscheinungsjahr

2022

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Endothelial cells line the lumen of all vessels in the body and maintain vascular homeostasis. In particular, endothelial cell regeneration in response to insult sustain functional endothelial layer. EdU (5-ethynyl-2′-deoxyuridine) is an alkyne-tagged proliferation probe that incorporates into newly synthesized DNA and is used for fluorescence imaging of cell proliferation with the use of “click chemistry” reaction with a fluorescent azide. Here, we utilized EdU as a click-free Raman probe for tracking endothelial cell proliferation. Raman imaging of EdU was performed in live endothelial cells, showing an advantage over fluorescence imaging of EdU, as this technique did not require sample fixation and permeabilization. To validate Raman-based imaging of EdU to study endothelial cell proliferation, we showed that when endothelial cells were treated with cycloheximide or doxorubicin to impair the proliferation of endothelial cells, the Raman-based signal of EdU was diminished. Furthermore, endothelial cells proliferation detected using EdU-labelled Raman imaging was compared with fluorescence imaging. Finally, the method of Raman–based EdU imaging was used in the isolated murine aorta ex vivo. Altogether, our results show that Raman-based imaging of EdU provides a novel alternative for fluorescence-based assay to assess endothelial proliferation and regeneration. •Raman probe of EdU molecule was used to follow endothelial cells (EC) proliferation in vitro and ex vivo.•Based on Raman imaging of EdU in a click-free manner it was possible to monitor DNA synthesis in cells.•The results of the EdU labelled Raman imaging to detect EC proliferation were confirmed using fluorescence microscopy.•Raman imaging of EdU provides a novel alternative for fluorescence-based endothelial proliferation and regeneration assays.