欧洲云杉针叶匀浆中的小细胞颗粒

Small Cellular Particles from European Spruce Needle Homogenate.

机构信息

University of Ljubljana, Laboratory of Clinical Biophysics, Faculty of Health Sciences, SI-1000 Ljubljana, Slovenia.

University of Ljubljana, Laboratory of Physics, Faculty of Electrical Engineering, SI-1000 Ljubljana, Slovenia.

出版信息

Int J Mol Sci. 2023 Feb 22;24(5):4349. doi: 10.3390/ijms24054349.

DOI:10.3390/ijms24054349

PMID:36901780

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10001569/

Abstract

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× 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× . The above results indicate that spruce needle homogenate is a source of vesicles to be explored for potential delivery use.

摘要

小细胞颗粒（SCPs）因其在细胞间通讯中的作用而受到关注。我们从云杉针叶匀浆中收获并表征了 SCPs。通过差速离心分离 SCPs。通过扫描电子显微镜（SEM）和低温透射电子显微镜（cryo-TEM）对其进行成像，通过干涉光显微镜（ILM）和流式细胞术（FCM）评估其数密度和水动力直径，通过紫外可见光谱法（UV-vis spectroscopy）测定总酚含量（TPC），通过气相色谱-质谱联用（GC-MS）测定萜烯含量。在 50,000×超速离心后的上清液中含有双层封闭囊泡，而在分离物中我们观察到其他类型的小颗粒，只有少数囊泡。细胞大小颗粒（CSPs）（大于 2 μm）和中大小颗粒（MSPs）（约 400 nm-2 μm）的数密度比 SCPs（小于 500 nm）的数密度低约四个数量级。在 10,029 个 SCPs 中测量的 SCPs 的平均水动力直径为 161 ± 133 nm。TCP 由于 5 天的老化而大大降低。在 300×离心后，在沉淀物中发现了挥发性萜类化合物。上述结果表明，云杉针叶匀浆是一种来源的囊泡，可用于探索潜在的递药用途。