Exosomes Produced from 3D Cultures of MSCs by Tangential Flow Filtration Show Higher Yield and Improved Activity

被引：463

作者：

Haraszti, Reka Agnes ^{[1
]}

Miller, Rachael ^{[3
]}

Stoppato, Matteo ^{[4
]}

Sere, Yves Y. ^{[4
]}

Coles, Andrew ^{[1
]}

Didiot, Marie-Cecile ^{[1
]}

Wollacott, Rachel ^{[4
]}

Sapp, Ellen ^{[5
]}

Dubuke, Michelle L. ^{[6
]}

Li, Xuni ^{[6
]}

Shaffer, Scott A. ^{[6
]}

DiFiglia, Marian ^{[5
]}

Wang, Yang ^{[4
]}

Aronin, Neil ^{[3
]}

Khvorova, Anastasia ^{[1
,2
]}

机构：

[1] Univ Massachusetts, Sch Med, RNA Therapeut Inst, 368 Plantat St, Worcester, MA 01605 USA

[2] Univ Massachusetts, Sch Med, Program Mol Med, Worcester, MA USA

[3] Univ Massachusetts, Sch Med, Dept Med, 368 Plantat St, Worcester, MA 01605 USA

[4] MassBiologics, Boston, MA USA

[5] Mass Gen Inst Neurodegenerat Dis, Boston, MA USA

[6] Univ Massachusetts, Sch Med, Mass Spectrometry Facil, Shrewsbury, MA USA

来源：

MOLECULAR THERAPY | 2018年 / 26卷 / 12期

关键词：

CELL-DERIVED EXOSOMES; HUNTINGTIN MESSENGER-RNA; EXTRACELLULAR VESICLES; MULTIVESICULAR BODIES; DENDRITIC CELLS; T-CELLS; DELIVERY; SIRNA; MICROVESICLES; PURIFICATION;

D O I：

10.1016/j.ymthe.2018.09.015

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 [微生物学]; 090105 [作物生产系统与生态工程];

摘要：

Exosomes can deliver therapeutic RNAs to neurons. The composition and the safety profile of exosomes depend on the type of the exosome-producing cell. Mesenchymal stem cells are considered to be an attractive cell type for therapeutic exosome production. However, scalable methods to isolate and manufacture exosomes from mesenchymal stem cells are lacking, a limitation to the clinical translation of exosome technology. We evaluate mesenchymal stem cells from different sources and find that umbilical cord-derived mesenchymal stem cells produce the highest exosome yield. To optimize exosome production, we cultivate umbilical cord-derived mesenchymal stem cells in scalable microcarrier-based three-dimensional (3D) cultures. In combination with the conventional differential ultracentrifugation, 3D culture yields 20-fold more exosomes (3D-UC-exosomes) than two-dimensional cultures (2D-UC-exosomes). Tangential flow filtration (TFF) in combination with 3D mesenchymal stem cell cultures further improves the yield of exosomes (3D-TFF-exosomes) 7-fold over 3D-UC-exosomes. 3D-TFF-exosomes are seven times more potent in small interfering RNA (siRNA) transfer to neurons compared with 2D-UC-exosomes. Microcarrier-based 3D culture and TFF allow scalable production of biologically active exosomes from mesenchymal stem cells. These findings lift a major roadblock for the clinical utility of mesenchymal stem cell exosomes.

引用

页码：2838 / 2847

页数：10

共 56 条

[1]

Hijacking Multivesicular Bodies Enables Long-Term and Exosome-Mediated Long-Distance Action of Anthrax Toxin [J].

Abrami, Laurence ;

Brandi, Lucia ;

Moayeri, Mahtab ;

Brown, Michael J. ;

Krantz, Bryan A. ;

Leppla, Stephen H. ;

van der Goot, F. Gisou .

CELL REPORTS, 2013, 5 (04) :986-996

[2]

Hydrophobically Modified siRNAs Silence Huntingtin mRNA in Primary Neurons and Mouse Brain [J].

Alterman, Julia F. ;

Hall, Lauren M. ;

Coles, Andrew H. ;

Hassler, Matthew R. ;

Didiot, Marie-Cecile ;

Chase, Kathryn ;

Abraham, Jasmin ;

Sottosanti, Emily ;

Johnson, Emily ;

Sapp, Ellen ;

Osborn, Maire F. ;

Difiglia, Marian ;

Aronin, Neil ;

Khvorova, Anastasia .

MOLECULAR THERAPY-NUCLEIC ACIDS, 2015, 4 :e266

[3]

Bone Marrow-Derived Mesenchymal Stromal Cells Harness Purinergenic Signaling to Tolerize Human Th1 Cells In Vivo [J].

Amarnath, Shoba ;

Foley, Jason E. ;

Farthing, Don E. ;

Gress, Ronald E. ;

Laurence, Arian ;

Eckhaus, Michael A. ;

Metais, Jean-Yves ;

Rose, Jeremy J. ;

Hakim, Frances T. ;

Felizardo, Tania C. ;

Cheng, Austin V. ;

Robey, Pamela G. ;

Stroncek, David E. ;

Sabatino, Marianna ;

Battiwalla, Minoo ;

Ito, Sawa ;

Fowler, Daniel H. ;

Barrett, Austin J. .

STEM CELLS, 2015, 33 (04) :1200-1212

[4]

Effects of Mesenchymal Stem Cell-Derived Exosomes on Experimental Autoimmune Uveitis [J].

Bai, Lingling ;

Shao, Hui ;

Wang, Hongxing ;

Zhang, Zhihui ;

Su, Chang ;

Dong, Lijie ;

Yu, Bo ;

Chen, Xiteng ;

Li, Xiaorong ;

Zhang, Xiaomin .

SCIENTIFIC REPORTS, 2017, 7

[5]

Human Pericardial Fluid Contains Exosomes Enriched with Cardiovascular-Expressed MicroRNAs and Promotes Therapeutic Angiogenesis [J].

Beltrami, Cristina ;

Besnier, Marie ;

Shantikumar, Saran ;

Shearn, Andrew I. U. ;

Rajakaruna, Cha ;

Laftah, Abas ;

Sessa, Fausto ;

Spinetti, Gaia ;

Petretto, Enrico ;

Angelini, Gianni D. ;

Emanueli, Costanza .

MOLECULAR THERAPY, 2017, 25 (03) :679-693

[6]

Manufacturing Exosomes: A Promising Therapeutic Platform [J].

Colao, Ivano Luigi ;

Corteling, Randolph ;

Bracewell, Daniel ;

Wall, Ivan .

TRENDS IN MOLECULAR MEDICINE, 2018, 24 (03) :242-256

[7]

A High-Throughput Method for Direct Detection of Therapeutic Oligonucleotide-Induced Gene Silencing In Vivo [J].

Coles, Andrew H. ;

Osborn, Maire F. ;

Alterman, Julia F. ;

Turanov, Anton A. ;

Godinho, Bruno M. D. C. ;

Kennington, Lori ;

Chase, Kathryn ;

Aronin, Neil ;

Khvorova, Anastasia .

NUCLEIC ACID THERAPEUTICS, 2016, 26 (02) :86-92

[8]

Reproducible and scalable purification of extracellular vesicles using combined bind-elute and size exclusion chromatography [J].

Corso, Giulia ;

Mager, Imre ;

Lee, Yi ;

Gorgens, Andre ;

Bultema, Jarred ;

Giebel, Bernd ;

Wood, Matthew J. A. ;

Nordin, Joel Z. ;

Andaloussi, Samir E. L. .

SCIENTIFIC REPORTS, 2017, 7

[9]

Exosome-mediated Delivery of Hydrophobically Modified siRNA for Huntingtin mRNA Silencing [J].

Didiot, Marie-Cecile ;

Hall, Lauren M. ;

Coles, Andrew H. ;

Haraszti, Reka A. ;

Godinho, Bruno M. D. C. ;

Chase, Kathryn ;

Sapp, Ellen ;

Ly, Socheata ;

Alterman, Julia F. ;

Hassler, Matthew R. ;

Echeverria, Dimas ;

Raj, Lakshmi ;

Morrissey, David V. ;

DiFiglia, Marian ;

Aronin, Neil ;

Khvorova, Anastasia .

MOLECULAR THERAPY, 2016, 24 (10) :1836-1847

[10]

Single pass tangential flow filtration to debottleneck downstream processing for therapeutic antibody production [J].

Dizon-Maspat, Jemelle ;

Bourret, Justin ;

D'Agostini, Anna ;

Li, Feng .

BIOTECHNOLOGY AND BIOENGINEERING, 2012, 109 (04) :962-970

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