성게 생식소 유래 세포외소포체 특성 분석 및 신경세포에 미치는 영향 연구
Characterization of Sea Urchin Gonad-derived Extracellular Vesicles and Study of Their Effects on Nerve Cells
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Extracellular vesicles (EVs) are nano-sized lipid bilayer vesicles released by cells. EVs act as messengers for cell-to-cell communication. Inside, it contains various substances that show biological activity, such as proteins, lipids, nucleic acids, and metabolites. The study of EVs extracted from terrestrial organisms and stem cells on inflam- matory environments and tissue regeneration have been actively conducted. However, marine organisms-derived EVs are limited. Therefore, we have extracted EVs from sea urchins belonging to the Echinoderm group with their excellent regenerative ability. First, we extracted extracellular matrix (ECM) from sea urchin gonads treated with hypotonic buffer, followed by collagenase treatment, and filtration to collect ECM-bounded EVs. The size of sea ur- chin gonad-derived EVs (UGEVs) is about 20-100 nm and has a round shape. The protein content was higher after EVs burst than before, which is evidence that proteins are contained inside. In addition, proteins of various sizes are distributed inside. PKH-26 was combined with UGEVs, which means that UGEVs have a lipid membrane. PHK-26- labeled UGEVs were successfully uptaken by cells. UGEVs can be confirmed to have the same characteristics as tra- ditional EVs. Finally, it was confirmed that Schwann cells were not toxic by increasing proliferation after treatment.
Abstract
Extracellular vesicles (EVs) are nano-sized lipid bilayer vesicles released by cells. EVs act as messengers for cell-to-cell communication. Inside, it contains various substances that show biological activity, such as proteins, lipids, nucleic acids, and metabolites. The study of EVs extracted from terrestrial organisms and stem cells on inflam- matory environments and tissue regeneration have been actively conducted. However, marine organisms-derived EVs are limited. Therefore, we have extracted EVs from sea urchins belonging to the Echinoderm group with their excellent regenerative ability. First, we extracted extracellular matrix (ECM) from sea urchin gonads treated with hypotonic buffer, followed by collagenase treatment, and filtration to collect ECM-bounded EVs. The size of sea ur- chin gonad-derived EVs (UGEVs) is about 20-100 nm and has a round shape. The protein content was higher after EVs burst than before, which is evidence that proteins are contained inside. In addition, proteins of various sizes are distributed inside. PKH-26 was combined with UGEVs, which means that UGEVs have a lipid membrane. PHK-26- labeled UGEVs were successfully uptaken by cells. UGEVs can be confirmed to have the same characteristics as tra- ditional EVs. Finally, it was confirmed that Schwann cells were not toxic by increasing proliferation after treatment.
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