Engineered Extracellular Vesicles and Their Mimetics for Clinical Translation.

Cells secrete extracellular vesicles (EVs) to external environments to achieve cellular homeostasis and cell-to-cell communication. Their therapeutic potential has been constantly spotlighted since they mirror both cytoplasmic and membranous components of parental cells. Meanwhile, growing evidence suggests that EV engineering could further promote EVs with a maximized capacity. In this review, a range of engineering techniques as well as upscaling approaches to exploit EVs and their mimetics are introduced. By laying out the pros and cons of each technique from different perspectives, we sought to provide an overview potentially helpful for understanding the current state of the art EV engineering and a guideline for choosing a suitable technique for engineering EVs. Furthermore, we envision that the advances in each technique will give rise to the combinatorial engineering of EVs, taking us a step closer to a clinical translation of EV-based therapeutics.

[1]  T. Anchordoquy,et al.  Biodistribution and delivery efficiency of unmodified tumor-derived exosomes. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[2]  Carmen Pazos,et al.  Therapeutic biomaterials based on extracellular vesicles: classification of bio-engineering and mimetic preparation routes , 2018, Journal of extracellular vesicles.

[3]  Michelle E. Hung,et al.  Stabilization of Exosome-targeting Peptides via Engineered Glycosylation* , 2015, The Journal of Biological Chemistry.

[4]  Dong Hwee Kim,et al.  Exosome as a Vehicle for Delivery of Membrane Protein Therapeutics, PH20, for Enhanced Tumor Penetration and Antitumor Efficacy , 2018 .

[5]  M. DiFiglia,et al.  Exosomes Produced from 3D Cultures of MSCs by Tangential Flow Filtration Show Higher Yield and Improved Activity. , 2018, Molecular therapy : the journal of the American Society of Gene Therapy.

[6]  Zhenjia Wang,et al.  Cell membrane-formed nanovesicles for disease-targeted delivery. , 2016, Journal of controlled release : official journal of the Controlled Release Society.

[7]  David Tareste,et al.  Modification of Extracellular Vesicles by Fusion with Liposomes for the Design of Personalized Biogenic Drug Delivery Systems. , 2018, ACS nano.

[8]  P. Robbins,et al.  Regulation of immune responses by extracellular vesicles , 2014, Nature Reviews Immunology.

[9]  Jian Song,et al.  A doxorubicin delivery platform using engineered natural membrane vesicle exosomes for targeted tumor therapy. , 2014, Biomaterials.

[10]  Ronnie H. Fang,et al.  Nanoparticulate Delivery of Cancer Cell Membrane Elicits Multiantigenic Antitumor Immunity , 2017, Advanced materials.

[11]  Heikki Saari,et al.  Microvesicle- and exosome-mediated drug delivery enhances the cytotoxicity of Paclitaxel in autologous prostate cancer cells. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[12]  L. O’Driscoll,et al.  Biological properties of extracellular vesicles and their physiological functions , 2015, Journal of extracellular vesicles.

[13]  R. Schiffelers,et al.  Recombinant phosphatidylserine-binding nanobodies for targeting of extracellular vesicles to tumor cells: a plug-and-play approach† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c7nr06966a , 2018, Nanoscale.

[14]  Yaojiong Wu,et al.  Three‐Dimensional Culture Reduces Cell Size By Increasing Vesicle Excretion , 2017, Stem cells.

[15]  S. Jay,et al.  Oncogene Knockdown via Active Loading of Small RNAs into Extracellular Vesicles by Sonication , 2016, Cellular and molecular bioengineering.

[16]  L. Tei,et al.  Efficient Route to Label Mesenchymal Stromal Cell-Derived Extracellular Vesicles , 2018, ACS omega.

[17]  S. Lim,et al.  Mesenchymal stem cell: an efficient mass producer of exosomes for drug delivery. , 2013, Advanced drug delivery reviews.

[18]  Jennifer C. Jones,et al.  Scalable, cGMP-compatible purification of extracellular vesicles carrying bioactive human heterodimeric IL-15/lactadherin complexes , 2018, Journal of extracellular vesicles.

[19]  Haifeng Dong,et al.  Engineered Exosome-Mediated Near-Infrared-II Region V2C Quantum Dot Delivery for Nucleus-Target Low-Temperature Photothermal Therapy. , 2019, ACS nano.

[20]  Alain Wagner,et al.  Copper-chelating azides for efficient click conjugation reactions in complex media. , 2014, Angewandte Chemie.

[21]  C. Barner‐Kowollik,et al.  Ultrafast click conjugation of macromolecular building blocks at ambient temperature. , 2009, Angewandte Chemie.

[22]  G. Dubyak,et al.  P2X7 receptors regulate multiple types of membrane trafficking responses and non-classical secretion pathways , 2009, Purinergic Signalling.

[23]  H. T. Park,et al.  RNAi delivery by exosome-mimetic nanovesicles - Implications for targeting c-Myc in cancer. , 2016, Biomaterials.

[24]  Molly M Stevens,et al.  Cell-derived vesicles for drug therapy and diagnostics: opportunities and challenges. , 2015, Nano today.

[25]  C. Mirkin,et al.  Exosome encased spherical nucleic acid gold nanoparticle conjugates as potent microRNA regulation agents. , 2014, Small.

[26]  José A López,et al.  Tissue-factor-bearing microvesicles arise from lipid rafts and fuse with activated platelets to initiate coagulation. , 2005, Blood.

[27]  John P. Fisher,et al.  Towards rationally designed biomanufacturing of therapeutic extracellular vesicles: impact of the bioproduction microenvironment. , 2018, Biotechnology advances.

[28]  G. Szabo,et al.  Extracellular vesicles in liver disease and potential as biomarkers and therapeutic targets , 2017, Nature Reviews Gastroenterology &Hepatology.

[29]  Stefania Raimondo,et al.  Interleukin 3- receptor targeted exosomes inhibit in vitro and in vivo Chronic Myelogenous Leukemia cell growth , 2017, Theranostics.

[30]  F. Sánchez‐Madrid,et al.  Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs , 2013, Nature Communications.

[31]  M. Perretti,et al.  Cutting-Edge Analysis of Extracellular Microparticles using ImageStreamX Imaging Flow Cytometry , 2014, Scientific Reports.

[32]  S. Singer Intercellular communication and cell-cell adhesion. , 1992, Science.

[33]  Carolyn R. Bertozzi,et al.  Chemical Technologies for Probing Glycans , 2006, Cell.

[34]  S. Wuttke,et al.  Exosome-Coated Metal-Organic Framework Nanoparticles: An Efficient Drug Delivery Platform , 2017 .

[35]  M. Vidal,et al.  Exosome Release Is Regulated by a Calcium-dependent Mechanism in K562 Cells* , 2003, Journal of Biological Chemistry.

[36]  Dylan T Burnette,et al.  Reassessment of Exosome Composition , 2019, Cell.

[37]  E. Bieberich,et al.  Ceramide and Exosomes: A Novel Target in Cancer Biology and Therapy: CHAPTER FIVE , 2018, Advances in cancer research.

[38]  P. Zou,et al.  Serum deprivation elevates the levels of microvesicles with different size distributions and selectively enriched proteins in human myeloma cells in vitro , 2013, Acta Pharmacologica Sinica.

[39]  Shinobu Ueda,et al.  Systemically Injected Exosomes Targeted to EGFR Deliver Antitumor MicroRNA to Breast Cancer Cells. , 2013, Molecular therapy : the journal of the American Society of Gene Therapy.

[40]  W. Möbius,et al.  Regulation of exosome secretion by Rab35 and its GTPase-activating proteins TBC1D10A–C , 2010, The Journal of cell biology.

[41]  C. Théry Exosomes: secreted vesicles and intercellular communications , 2011, F1000 biology reports.

[42]  Alexander V Kabanov,et al.  TPP1 Delivery to Lysosomes with Extracellular Vesicles and their Enhanced Brain Distribution in the Animal Model of Batten Disease , 2019, Advanced healthcare materials.

[43]  Imre Mäger,et al.  Extracellular vesicles: biology and emerging therapeutic opportunities , 2013, Nature Reviews Drug Discovery.

[44]  I. Sargent,et al.  Exosome-mediated delivery of siRNA in vitro and in vivo , 2012, Nature Protocols.

[45]  Jing Xu,et al.  Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines , 2018, Journal of Extracellular Vesicles.

[46]  M. Ferguson,et al.  Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. , 1988, Annual review of biochemistry.

[47]  In‐San Kim,et al.  Extracellular vesicles as a platform for membrane-associated therapeutic protein delivery , 2018, Journal of extracellular vesicles.

[48]  Guodong Yang,et al.  In Vitro and in Vivo RNA Inhibition by CD9-HuR Functionalized Exosomes Encapsulated with miRNA or CRISPR/dCas9. , 2018, Nano letters.

[49]  James P K Armstrong,et al.  Re-Engineering Extracellular Vesicles as Smart Nanoscale Therapeutics. , 2017, ACS nano.

[50]  Weian Zhao,et al.  Stem Cell Extracellular Vesicles: Extended Messages of Regeneration , 2016, Annual review of pharmacology and toxicology.

[51]  Jaesung Park,et al.  Bioinspired exosome-mimetic nanovesicles for targeted delivery of chemotherapeutics to malignant tumors. , 2013, ACS nano.

[52]  Molly M Stevens,et al.  Active loading into extracellular vesicles significantly improves the cellular uptake and photodynamic effect of porphyrins. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[53]  Christopher H Contag,et al.  Differential fates of biomolecules delivered to target cells via extracellular vesicles , 2015, Proceedings of the National Academy of Sciences.

[54]  Simon C Watkins,et al.  Exosomes Derived from IL-10-Treated Dendritic Cells Can Suppress Inflammation and Collagen-Induced Arthritis 1 , 2005, The Journal of Immunology.

[55]  Gong Cheng,et al.  Aptamer-Conjugated Extracellular Nanovesicles for Targeted Drug Delivery. , 2018, Cancer research.

[56]  M. Pittenger,et al.  Concise Review: MSC‐Derived Exosomes for Cell‐Free Therapy , 2017, Stem cells.

[57]  Xiaolong Liu,et al.  Chemotherapeutic Drug Based Metal–Organic Particles for Microvesicle‐Mediated Deep Penetration and Programmable pH/NIR/Hypoxia Activated Cancer Photochemotherapy , 2018, Advanced science.

[58]  Ivan Wall,et al.  Manufacturing Exosomes: A Promising Therapeutic Platform. , 2018, Trends in molecular medicine.

[59]  Laurence Zitvogel,et al.  Exosomes: composition, biogenesis and function , 2002, Nature Reviews Immunology.

[60]  Per Sunnerhagen,et al.  Plasma exosomes can deliver exogenous short interfering RNA to monocytes and lymphocytes , 2012, Nucleic acids research.

[61]  M. Record,et al.  PLD2 is enriched on exosomes and its activity is correlated to the release of exosomes , 2004, FEBS letters.

[62]  Ronnie H. Fang,et al.  Nanoparticle biointerfacing via platelet membrane cloaking , 2015, Nature.

[63]  Qiaobing Xu,et al.  Integrating Protein Engineering and Bioorthogonal Click Conjugation for Extracellular Vesicle Modulation and Intracellular Delivery , 2015, PloS one.

[64]  Liesbet Geris,et al.  Towards Self-Regulated Bioprocessing: A Compact Benchtop Bioreactor System for Monitored and Controlled 3D Cell and Tissue Culture. , 2019, Biotechnology journal.

[65]  S. Gambhir,et al.  Tumor Cell-Derived Extracellular Vesicle-Coated Nanocarriers: An Efficient Theranostic Platform for the Cancer-Specific Delivery of Anti-miR-21 and Imaging Agents. , 2018, ACS nano.

[66]  Thomas J. Anchordoquy,et al.  Surface Functionalization of Exosomes Using Click Chemistry , 2014, Bioconjugate chemistry.

[67]  Ylva Ivarsson,et al.  Syndecan–syntenin–ALIX regulates the biogenesis of exosomes , 2012, Nature Cell Biology.

[68]  Y. Weizmann,et al.  Mesenchymal stem cell and derived exosome as small RNA carrier and Immunomodulator to improve islet transplantation. , 2016, Journal of controlled release : official journal of the Controlled Release Society.

[69]  Song Fan,et al.  Surface functionalized exosomes as targeted drug delivery vehicles for cerebral ischemia therapy. , 2018, Biomaterials.

[70]  C. Oliver,et al.  Permeabilization of cell membranes. , 2010, Methods in molecular biology.

[71]  A. Weisz,et al.  The RNA-Binding Protein SYNCRIP Is a Component of the Hepatocyte Exosomal Machinery Controlling MicroRNA Sorting. , 2016, Cell reports.

[72]  Y. Takakura,et al.  Exosome-based tumor antigens-adjuvant co-delivery utilizing genetically engineered tumor cell-derived exosomes with immunostimulatory CpG DNA. , 2016, Biomaterials.

[73]  C. Wilhelm,et al.  Extracellular vesicles for personalized medicine: The input of physically triggered production, loading and theranostic properties. , 2019, Advanced drug delivery reviews.

[74]  K. Braeckmans,et al.  Electroporation-induced siRNA precipitation obscures the efficiency of siRNA loading into extracellular vesicles. , 2013, Journal of controlled release : official journal of the Controlled Release Society.

[75]  J. Lötvall,et al.  Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells , 2007, Nature Cell Biology.

[76]  Maria Carmen Blanco-López,et al.  Fully Artificial Exosomes: Towards New Theranostic Biomaterials. , 2018, Trends in biotechnology.

[77]  Zhiping Zhang,et al.  Extracellular vesicles based self-grown gold nanopopcorn for combinatorial chemo-photothermal therapy. , 2019, Biomaterials.

[78]  Thomas D. Schmittgen,et al.  Low active loading of cargo into engineered extracellular vesicles results in inefficient miRNA mimic delivery , 2017, Journal of extracellular vesicles.

[79]  R. Schiffelers,et al.  PEGylated and targeted extracellular vesicles display enhanced cell specificity and circulation time. , 2016, Journal of controlled release : official journal of the Controlled Release Society.

[80]  Sanchita Bhatnagar,et al.  Exosome Function: From Tumor Immunology to Pathogen Biology , 2008, Traffic.

[81]  Julia Christina Gross,et al.  Active Wnt proteins are secreted on exosomes , 2012, Nature Cell Biology.

[82]  M. Wood,et al.  Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes , 2011, Nature Biotechnology.

[83]  Jinghuan Li,et al.  Serum-free culture alters the quantity and protein composition of neuroblastoma-derived extracellular vesicles , 2015, Journal of extracellular vesicles.

[84]  A. Llorente,et al.  Exosomal lipid composition and the role of ether lipids and phosphoinositides in exosome biology , 2018, Journal of Lipid Research.

[85]  Jennifer C Jones,et al.  Efficient production and enhanced tumor delivery of engineered extracellular vesicles. , 2016, Biomaterials.

[86]  F. Wendler,et al.  Extracellular vesicles round off communication in the nervous system , 2016, Nature Reviews Neuroscience.

[87]  U. Ruktanonchai,et al.  Surface modification of gold nanoparticles with neuron-targeted exosome for enhanced blood–brain barrier penetration , 2019, Scientific Reports.

[88]  Yanping Sun,et al.  Recent advances on extracellular vesicles in therapeutic delivery: Challenges, solutions, and opportunities , 2017, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[89]  Martin Fussenegger,et al.  Designer exosomes produced by implanted cells intracerebrally deliver therapeutic cargo for Parkinson’s disease treatment , 2018, Nature Communications.

[90]  Jacopo Meldolesi,et al.  Shedding microvesicles: artefacts no more. , 2009, Trends in cell biology.

[91]  G. Willis,et al.  Nitrite-derived nitric oxide reduces hypoxia-inducible factor 1α-mediated extracellular vesicle production by endothelial cells. , 2017, Nitric oxide : biology and chemistry.

[92]  M. Yáñez-Mó,et al.  Tetraspanins in Extracellular Vesicle Formation and Function , 2014, Front. Immunol..

[93]  S. Kourembanas,et al.  Toward Exosome-Based Therapeutics: Isolation, Heterogeneity, and Fit-for-Purpose Potency , 2017, Front. Cardiovasc. Med..

[94]  Peixuan Guo,et al.  Nanoparticle Orientation to Control RNA Loading and Ligand Display on Extracellular Vesicles for Cancer Regression , 2017, Nature Nanotechnology.

[95]  Ji-Ho Park,et al.  Cooperative tumour cell membrane targeted phototherapy , 2017, Nature Communications.

[96]  G. Pastorin,et al.  Doxorubicin-loaded cell-derived nanovesicles: an alternative targeted approach for anti-tumor therapy , 2017, International journal of nanomedicine.

[97]  Yong Song Gho,et al.  Importance of exosome depletion protocols to eliminate functional and RNA-containing extracellular vesicles from fetal bovine serum , 2014, Journal of extracellular vesicles.

[98]  A. Sinz,et al.  Chances and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters , 2008, Analytical and bioanalytical chemistry.

[99]  E. Clementi,et al.  Acid sphingomyelinase activity triggers microparticle release from glial cells , 2009, The EMBO journal.

[100]  Graça Raposo,et al.  Extracellular vesicles: Exosomes, microvesicles, and friends , 2013, The Journal of cell biology.

[101]  Andrew L. Ferguson,et al.  Polymeric "Clickase" Accelerates the Copper Click Reaction of Small Molecules, Proteins, and Cells. , 2019, Journal of the American Chemical Society.

[102]  Eun Hee Kim,et al.  Efficient scalable production of therapeutic microvesicles derived from human mesenchymal stem cells , 2018, Scientific Reports.

[103]  Phillip C. Yang,et al.  Exosomes Generated From iPSC-Derivatives: New Direction for Stem Cell Therapy in Human Heart Diseases , 2017, Circulation research.

[104]  Jennifer A. Prescher,et al.  Copper-free click chemistry in living animals , 2010, Proceedings of the National Academy of Sciences.

[105]  Miguel C. Seabra,et al.  Rab27a and Rab27b control different steps of the exosome secretion pathway , 2010, Nature Cell Biology.

[106]  N. Vitale,et al.  Syntenin-ALIX exosome biogenesis and budding into multivesicular bodies are controlled by ARF6 and PLD2 , 2014, Nature Communications.

[107]  Pieter Vader,et al.  Extracellular vesicles for drug delivery. , 2016, Advanced drug delivery reviews.

[108]  Clotilde Théry,et al.  Communication by Extracellular Vesicles: Where We Are and Where We Need to Go , 2016, Cell.

[109]  S. Tenzer,et al.  Oligodendrocytes secrete exosomes containing major myelin and stress‐protective proteins: Trophic support for axons? , 2007, Proteomics. Clinical applications.

[110]  D. Pang,et al.  Transformation of cell-derived microparticles into quantum-dot-labeled nanovectors for antitumor siRNA delivery. , 2015, Angewandte Chemie.

[111]  Qingfu Zhu,et al.  Microfluidic engineering of exosomes: editing cellular messages for precision therapeutics. , 2018, Lab on a chip.

[112]  Xiaojing Liu,et al.  Aptamer-Functionalized Exosomes: Elucidating the Cellular Uptake Mechanism and the Potential for Cancer-Targeted Chemotherapy. , 2019, Analytical chemistry.

[113]  C. Tung,et al.  Facile metabolic glycan labeling strategy for exosome tracking. , 2018, Biochimica et biophysica acta. General subjects.

[114]  Ji-Ho Park,et al.  Cellular Engineering with Membrane Fusogenic Liposomes to Produce Functionalized Extracellular Vesicles. , 2016, ACS applied materials & interfaces.

[115]  Ji-Ho Park,et al.  Liposome-based engineering of cells to package hydrophobic compounds in membrane vesicles for tumor penetration. , 2015, Nano letters.

[116]  Luigi Biancone,et al.  Exosomes/microvesicles as a mechanism of cell-to-cell communication. , 2010, Kidney international.

[117]  Steven M Jay,et al.  Exogenous DNA Loading into Extracellular Vesicles via Electroporation is Size-Dependent and Enables Limited Gene Delivery. , 2015, Molecular pharmaceutics.

[118]  M. Stevens,et al.  Engineering Extracellular Vesicles with the Tools of Enzyme Prodrug Therapy , 2018, Advanced materials.

[119]  Myung Soo Kim,et al.  Engineering macrophage-derived exosomes for targeted paclitaxel delivery to pulmonary metastases: in vitro and in vivo evaluations. , 2018, Nanomedicine : nanotechnology, biology, and medicine.

[120]  Antonella Bongiovanni,et al.  Scalable Production and Isolation of Extracellular Vesicles: Available Sources and Lessons from Current Industrial Bioprocesses , 2019, Biotechnology journal.

[121]  Yoosoo Yang,et al.  Exosome-SIRPα, a CD47 blockade increases cancer cell phagocytosis. , 2017, Biomaterials.

[122]  Kwang Ryeol Lee,et al.  Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein–protein interaction module , 2016, Nature Communications.

[123]  N. Gu,et al.  Light‐Inducible Exosome‐Based Vehicle for Endogenous RNA Loading and Delivery to Leukemia Cells , 2019, Advanced Functional Materials.

[124]  Shunichi Homma,et al.  Cardiac recovery via extended cell-free delivery of extracellular vesicles secreted by cardiomyocytes derived from induced pluripotent stem cells , 2018, Nature Biomedical Engineering.

[125]  Petra Schwille,et al.  Ceramide Triggers Budding of Exosome Vesicles into Multivesicular Endosomes , 2008, Science.

[126]  Chang-Qing Zhang,et al.  Exosomes derived from miR-140-5p-overexpressing human synovial mesenchymal stem cells enhance cartilage tissue regeneration and prevent osteoarthritis of the knee in a rat model , 2017, Theranostics.

[127]  P. Paz,et al.  Exosome Display technology: applications to the development of new diagnostics and therapeutics. , 2005, Blood cells, molecules & diseases.

[128]  J Ratajczak,et al.  Embryonic stem cell-derived microvesicles reprogram hematopoietic progenitors: evidence for horizontal transfer of mRNA and protein delivery , 2006, Leukemia.

[129]  C. Théry,et al.  Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. , 2014, Annual review of cell and developmental biology.

[130]  Graca Raposo,et al.  ARF6-Regulated Shedding of Tumor Cell-Derived Plasma Membrane Microvesicles , 2009, Current Biology.

[131]  Z. Madeja,et al.  Human Induced Pluripotent Stem Cell‐Derived Microvesicles Transmit RNAs and Proteins to Recipient Mature Heart Cells Modulating Cell Fate and Behavior , 2015, Stem cells.

[132]  Wei Zhang,et al.  Magnetic and Folate Functionalization Enables Rapid Isolation and Enhanced Tumor-Targeting of Cell-Derived Microvesicles. , 2017, ACS nano.

[133]  A. Mikos,et al.  Flow perfusion effects on three-dimensional culture and drug sensitivity of Ewing sarcoma , 2015, Proceedings of the National Academy of Sciences.

[134]  V. Lee,et al.  A "Clickable" Photoconvertible Small Fluorescent Molecule as a Minimalist Probe for Tracking Individual Biomolecule Complexes. , 2019, Journal of the American Chemical Society.

[135]  Graça Raposo,et al.  Shedding light on the cell biology of extracellular vesicles , 2018, Nature Reviews Molecular Cell Biology.

[136]  Jacco van Rheenen,et al.  In Vivo Imaging Reveals Extracellular Vesicle-Mediated Phenocopying of Metastatic Behavior , 2015, Cell.

[137]  M. Epple,et al.  Covalent Surface Functionalization of Calcium Phosphate Nanoparticles with Fluorescent Dyes by Copper‐Catalysed and by Strain‐Promoted Azide‐Alkyne Click Chemistry , 2018, ChemNanoMat.

[138]  J. Fisher,et al.  Enhanced extracellular vesicle production and ethanol-mediated vascularization bioactivity via a 3D-printed scaffold-perfusion bioreactor system. , 2019, Acta biomaterialia.

[139]  Richa Gupta,et al.  Exosomes as drug delivery vehicles for Parkinson's disease therapy. , 2015, Journal of controlled release : official journal of the Controlled Release Society.

[140]  C. Lehr,et al.  Extracellular vesicles protect glucuronidase model enzymes during freeze-drying , 2018, Scientific Reports.

[141]  Myung Soo Kim,et al.  Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells. , 2016, Nanomedicine : nanotechnology, biology, and medicine.

[142]  Pieter Vader,et al.  Display of GPI-anchored anti-EGFR nanobodies on extracellular vesicles promotes tumour cell targeting , 2016, Journal of extracellular vesicles.