Efficient scalable production of therapeutic microvesicles derived from human mesenchymal stem cells

Jae Min Cha,Gyeong Joon Moon,Ji Hee Sung,Hyung Dal Park,Eun Hee Kim,Eun Kyoung Shin,Jinseok Kim,Hojae Bae,Yeon Hee Cho,Oh Young Bang

doi:10.1038/s41598-018-19211-6

Abstract

Microvesicles (MVs) released by cells are involved in a multitude of physiological events as important mediators of intercellular communication. MVs derived from mesenchymal stem cells (MSCs) contain various paracrine factors from the cells that primarily contribute to their therapeutic efficacy observed in numerous clinical trials. As nano-sized and bi-lipid layered vesicles retaining therapeutic potency equivalent to that of MSCs, MSC-derived MVs have been in focus as ideal medicinal candidates for regenerative medicine, and are preferred over MSC infusion therapy with their improved safety profiles. However, technical challenges in obtaining sufficient amounts of MVs have limited further progress in studies and clinical application. Of the multiple efforts to reinforce the therapeutic capacity of MSCs, few studies have reportedly examined the scale-up of MSC-derived MV production. In this study, we successfully amplified MV secretion from MSCs compared to the conventional culture method using a simple and efficient 3D-bioprocessing method. The MSC-derived MVs produced in our dynamic 3D-culture contained numerous therapeutic factors such as cytokines and micro-RNAs, and showed their therapeutic potency in in vitro efficacy evaluation. Our results may facilitate diverse applications of MSC-derived MVs from the bench to the bedside, which requires the large-scale production of MVs.

Highlights

Numerous clinical trials of mesenchymal stem cells (MSCs) are being conducted to develop effective treatments for diseases with few curable options, such as stroke, spinal cord injury, multiple sclerosis, Alzheimer’s disease, liver cirrhosis, myocardial infarction, kidney disease, and graft-versus-host disease, among others
In addition to the dynamic 3D-human MSC (hMSC) culture employed in this study, we previously reported that the efficacy of hMSCs could be enhanced by pretreating the culture with ischemic brain extract (IBE)[33]
Extracellular vesicles (EVs) released by cells play a crucial role in regulating numerous physiological events and orchestrating interactions between cells and the surrounding microenvironments[30]

Summary

Introduction

Numerous clinical trials of mesenchymal stem cells (MSCs) are being conducted to develop effective treatments for diseases with few curable options, such as stroke, spinal cord injury, multiple sclerosis, Alzheimer’s disease, liver cirrhosis, myocardial infarction, kidney disease, and graft-versus-host disease, among others. (www.clinicaltrials.gov). As reported by numerous previous studies, the intrinsic ability of MSCs to secrete a variety of therapeutic molecules is difficult to reproduce in vitro, as natural 3D-interactions between cells and either the extracellular matrix (ECM) or other neighbouring cells are readily disrupted under conventional monolayer conditions where individual cells encounter limited two-dimensional borders[16,17]. We successfully amplified therapeutic MSC-derived MVs by using a simple and effectual dynamic 3D-culture method (Fig. 1c). By subsequently providing a dynamic culture environment, the production of MSC-derived MVs was significantly increased compared to that using a conventional monolayer culture method. The inclusion of a variety of therapeutic factors in MSC-derived MVs from our dynamic 3D-culture was investigated, and their medicinal potency was evaluated using different culture models

Methods

Results

Conclusion