Small Cellular Particles from European Spruce Needle Homogenate
Small cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmiss...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/5/4349 |
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| author | Marko Jeran Anna Romolo Vesna Spasovski Matej Hočevar Urban Novak Roman Štukelj Vid Šuštar Matic Kisovec Apolonija Bedina Zavec Ksenija Kogej Aleš Iglič Polonca Trebše Veronika Kralj-Iglič |
| author_facet | Marko Jeran Anna Romolo Vesna Spasovski Matej Hočevar Urban Novak Roman Štukelj Vid Šuštar Matic Kisovec Apolonija Bedina Zavec Ksenija Kogej Aleš Iglič Polonca Trebše Veronika Kralj-Iglič |
| author_sort | Marko Jeran |
| collection | DOAJ |
| description | Small cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmission electron microscope (cryo TEM), assessed for their number density and hydrodynamic diameter by interferometric light microscopy (ILM) and flow cytometry (FCM), total phenolic content (TPC) by UV-vis spectroscopy, and terpene content by gas chromatography-mass spectrometry (GC-MS). The supernatant after ultracentrifugation at 50,000× <i>g</i> contained bilayer-enclosed vesicles whereas in the isolate we observed small particles of other types and only a few vesicles. The number density of cell-sized particles (CSPs) (larger than 2 μm) and meso-sized particles (MSPs) (cca 400 nm–2 µm) was about four orders of magnitude lower than the number density of SCPs (sized below 500 nm). The average hydrodynamic diameter of SCPs measured in 10,029 SCPs was 161 ± 133 nm. TCP decreased considerably due to 5-day aging. Volatile terpenoid content was found in the pellet after 300× <i>g</i>. The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use. |
| first_indexed | 2024-03-11T07:23:35Z |
| format | Article |
| id | doaj.art-6b72cd7c2e1f4aef8a13292b7ff89c67 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T07:23:35Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-6b72cd7c2e1f4aef8a13292b7ff89c672023-11-17T07:47:14ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01245434910.3390/ijms24054349Small Cellular Particles from European Spruce Needle HomogenateMarko Jeran0Anna Romolo1Vesna Spasovski2Matej Hočevar3Urban Novak4Roman Štukelj5Vid Šuštar6Matic Kisovec7Apolonija Bedina Zavec8Ksenija Kogej9Aleš Iglič10Polonca Trebše11Veronika Kralj-Iglič12University of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaInstitute for Metals and Technology, SI-1000 Ljubljana, SloveniaTheory Department, National Institute of Chemistry, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaDepartment of Molecular Biology and Nanobiotechnology, National Institute of Chemistry, SI-1000 Ljubljana, SloveniaDepartment of Molecular Biology and Nanobiotechnology, National Institute of Chemistry, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Faculty of Chemistry and Chemical Technology, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Physics, Faculty of Electrical Engineering, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaUniversity of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, SloveniaSmall cellular particles (SCPs) are being considered for their role in cell-to-cell communication. We harvested and characterized SCPs from spruce needle homogenate. SCPs were isolated by differential ultracentrifugation. They were imaged by scanning electron microscope (SEM) and cryogenic transmission electron microscope (cryo TEM), assessed for their number density and hydrodynamic diameter by interferometric light microscopy (ILM) and flow cytometry (FCM), total phenolic content (TPC) by UV-vis spectroscopy, and terpene content by gas chromatography-mass spectrometry (GC-MS). The supernatant after ultracentrifugation at 50,000× <i>g</i> contained bilayer-enclosed vesicles whereas in the isolate we observed small particles of other types and only a few vesicles. The number density of cell-sized particles (CSPs) (larger than 2 μm) and meso-sized particles (MSPs) (cca 400 nm–2 µm) was about four orders of magnitude lower than the number density of SCPs (sized below 500 nm). The average hydrodynamic diameter of SCPs measured in 10,029 SCPs was 161 ± 133 nm. TCP decreased considerably due to 5-day aging. Volatile terpenoid content was found in the pellet after 300× <i>g</i>. The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use.https://www.mdpi.com/1422-0067/24/5/4349<i>Picea abies</i>spruce needlesextracellular particlesextracellular vesiclessmall cellular particlesinterferometric light microscopy |
| spellingShingle | Marko Jeran Anna Romolo Vesna Spasovski Matej Hočevar Urban Novak Roman Štukelj Vid Šuštar Matic Kisovec Apolonija Bedina Zavec Ksenija Kogej Aleš Iglič Polonca Trebše Veronika Kralj-Iglič Small Cellular Particles from European Spruce Needle Homogenate International Journal of Molecular Sciences <i>Picea abies</i> spruce needles extracellular particles extracellular vesicles small cellular particles interferometric light microscopy |
| title | Small Cellular Particles from European Spruce Needle Homogenate |
| title_full | Small Cellular Particles from European Spruce Needle Homogenate |
| title_fullStr | Small Cellular Particles from European Spruce Needle Homogenate |
| title_full_unstemmed | Small Cellular Particles from European Spruce Needle Homogenate |
| title_short | Small Cellular Particles from European Spruce Needle Homogenate |
| title_sort | small cellular particles from european spruce needle homogenate |
| topic | <i>Picea abies</i> spruce needles extracellular particles extracellular vesicles small cellular particles interferometric light microscopy |
| url | https://www.mdpi.com/1422-0067/24/5/4349 |
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