Sharma Shayna, Scholz-Romero Katherin, Rice Gregory E, Salomon Carlos
Exosome Biology Laboratory, Centre for Clinical Diagnostics, University of Queensland Centre for Clinical Research, Royal Brisbane and Women's Hospital, The University of Queensland, Bldg. 71/918, Royal Brisbane Hospital, Brisbane, QLD, 4029, Australia.
Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Ochsner Clinic Foundation, New Orleans, LA, USA.
Methods Mol Biol. 2018;1710:103-115. doi: 10.1007/978-1-4939-7498-6_8.
Exosomes are nano-vesicles which can transport a range of molecules including but not limited to proteins and miRNA. This ability of exosomes renders them useful in cellular communication often resulting in biological changes. They have several functions in facilitating normal biological processes such as immune responses and an involvement in pregnancy. However, they have also been linked to pathological conditions including cancer and pregnancy complications such as preeclampsia. An understanding for the role of exosomes in preeclampsia is based on the ability to purify and characterize exosomes. There have been several techniques proposed for the enrichment of exosomes such as ultracentrifugation, density gradient separation, and ultrafiltration although there is no widely accepted optimized technique. Here we describe a workflow for isolating exosomes from cell-conditioned media and biological fluids using a combination of centrifugation, buoyant density, and ultrafiltration approaches.
外泌体是纳米囊泡，能够运输一系列分子，包括但不限于蛋白质和微小RNA。外泌体的这种能力使其在细胞通讯中发挥作用，常常导致生物学变化。它们在促进正常生物学过程（如免疫反应）以及参与妊娠方面具有多种功能。然而，它们也与包括癌症在内的病理状况以及诸如子痫前期等妊娠并发症有关。对外泌体在子痫前期中作用的理解基于纯化和表征外泌体的能力。虽然目前尚无广泛接受的优化技术，但已经提出了几种用于富集外泌体的技术，如超速离心、密度梯度分离和超滤。在此，我们描述了一种使用离心、浮力密度和超滤方法相结合，从细胞条件培养基和生物流体中分离外泌体的工作流程。
