Cell-engineered nanovesicles (CNVs) are considered as an alternative to exosomes, because they can be produced efficiently on a large scale and have been successfully reported in several applied research studies. However, CNVs may originate from various organelles, i.e., some of them may cause adverse effects on recipient cells, and their origin has not yet been identified. In this study, we air-sprayed human embryonic kidney 293 (HEK293) cells into lipid-bilayer CNVs. To identify the subcellular origin of the CNVs, we prepared nine different HEK293 cell lines by transfection with organelle-specific fluorescent protein plasmids that target the plasma membrane, peroxisome, lysosome, early endosome, late endosome, nucleus, mitochondrion, Golgi apparatus, and endoplasmic reticulum. The origin of CNVs were identified by measuring fluorescence expressions for organelle-specific markers using fluorescence nanoparticle tracking analysis (NTA). In the results, we found that CNVs derived from the plasma membrane constituted the largest portion, but CNVs derived from the other organelles comprised a non-negligible portion as well. This information will be useful to guide advanced research on outer membrane vesicles and exosome-mimetic nanovesicles engineered from cells.

中文翻译：

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从细胞工程设计的外泌体模拟纳米囊泡的异质亚细胞起源。

细胞工程化的纳米囊泡（CNV）被认为是外泌体的替代物，因为它们可以大规模高效生产，并已在一些应用研究中成功报道。但是，CNV可能起源于各种细胞器，即其中一些可能对受体细胞产生不利影响，其来源尚未确定。在这项研究中，我们将人类胚胎肾293（HEK293）细胞通过空气喷涂成脂质双层CNV。为了鉴定CNV的亚细胞起源，我们通过用细胞器特异性荧光蛋白质粒转染制备了9种不同的HEK293细胞系，这些质粒靶向质膜，过氧化物酶体，溶酶体，早期内体，晚期内体，细胞核，线粒体，高尔基体和内质网状。通过使用荧光纳米颗粒跟踪分析（NTA）测量细胞器特异性标记的荧光表达来鉴定CNV的起源。在结果中，我们发现衍生自质膜的CNV构成了最大部分，而衍生自其他细胞器的CNV也包含了不可忽略的部分。这些信息将有助于指导从细胞工程化的外膜囊泡和模拟外泌体纳米囊泡的高级研究。