National Institutes of Health (NIH), Ministry of Health Malaysia 1 article published in JoVE Biology A Simple Benchtop Filtration Method to Isolate Small Extracellular Vesicles from Human Mesenchymal Stem Cells Benson Koh1,2, Kian Leong Tan3, Hong Hao Chan4, Qi Hao Daniel Looi2, Moon Nian Lim5, Chee Wun How4, Jia Xian Law1, Jhi Biau Foo6,7 1Centre for Tissue Engineering and Regenerative Medicine, Universiti Kebangsaan Malaysia Medical Centre, 2My CytoHealth Sdn. Bhd., 3School of Pharmacy, Faculty of Health & Medical Sciences, Taylor's University, 7 This protocol demonstrates how to isolate human umbilical cord-derived mesenchymal stem cells' small extracellular vesicles (hUC-MSC-sEVs) with a simple lab-scale benchtop setting. The size distribution, protein concentration, sEVs markers, and morphology of isolated hUC-MSC-sEVs are characterized by nanoparticle tracking analysis, BCA protein assay, western blot, and transmission electron microscope, respectively.