Signal Factors Secreted by 2D and Spheroid Mesenchymal Stem Cells and by Cocultures of Mesenchymal Stem Cells Derived Microvesicles and Retinal Photoreceptor Neurons.

Lili Xie,Lusi Zhang,Liang Zhou,Bing Jiang,Mao Mao

doi:10.1155/2017/2730472

Abstract

We aim to identify levels of signal factors secreted by MSCs cultured in 2D monolayers (2D-MSCs), spheroids (spheroids MSCs), and cocultures of microvesicles (MVs) derived from 2D-MSCs or spheroid MSCs and retinal photoreceptor neurons. We seeded 2D-MSCs, spheroid MSCs, and cells derived from spheroids MSCs at equal numbers. MVs isolated from all 3 culture conditions were incubated with 661W cells. Levels of 51 signal factors in conditioned medium from those cultured conditions were quantified with bead-based assay. We found that IL-8, IL-6, and GROα were the top three most abundant signal factors. Moreover, compared to 2D-MSCs, levels of 11 cytokines and IL-2Rα were significantly increased in conditioned medium from spheroid MSCs. Finally, to test if enhanced expression of these factors reflects altered immunomodulating activities, we assessed the effect of 2D-MSC-MVs and 3D-MSC-MVs on CD14+ cell chemoattraction. Compared to 2D-MSC-MVs, 3D-MSC-MVs significantly decreased the chemotactic index of CD14+ cells. Our results suggest that spheroid culture conditions improve the ability of MSCs to selectively secrete signal factors. Moreover, 3D-MSC-MVs also possessed an enhanced capability to promote signal factors secretion compared to 2D-MSC-MVs and may possess enhanced immunomodulating activities and might be a better regenerative therapy for retinal degenerative diseases.

Highlights

There has been considerable interest in the curative effect of the human mesenchymal stem cells (MSCs) that are derived from adult tissues such as umbilical cord blood, bone marrow, and adipose tissue [1,2,3]
Our study aimed to systematically analyze signal factors secreted by 2D-MSCs and spheroid MSCs and the effect of 2D-MSC-MVs and 3D-MSC-MVs on signal factor secretion when cocultured with retinal photoreceptor neurons
IL-8, IL-6, and GROα Are the Top Three Cytokines in Concentration Secreted by 2D-MSCs

Summary

Introduction

There has been considerable interest in the curative effect of the human mesenchymal stem cells (MSCs) that are derived from adult tissues such as umbilical cord blood, bone marrow, and adipose tissue [1,2,3]. MSCs are relatively convenient to be isolated from donors, and they can maintain an active proliferating capacity after multiple passages in culture. For these reasons, MSCs have great therapeutic potential in disease treatment, as demonstrated by results from multiple experimental and clinical studies [4,5,6]. MSCs have great therapeutic potential in disease treatment, as demonstrated by results from multiple experimental and clinical studies [4,5,6] In addition to their multidifferentiation potential, MSCs are well known for their abilities to secrete paracrine factors and to modulate inflammation and immunity [7, 8]. Increasing attention has been paid to ways to enhance MSC treatment efficiency and to identify MSCderived elements conferring potent neuroprotection [14]

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