Regenerative efficacy of mesenchymal stromal cells from human placenta in sensorineural hearing loss

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Abstract

Introduction

Hearing loss is a common chronic disorder characterized by decline of auditory function. The global population have suffered from deafness and the transplantation of stem cells is regarded as a therapeutic strategy for this disease.

Methods

We collected placenta from a total of 13 samples of full term pregnant women and isolated MSCs derived from human placenta and transplanted MSCs on deaf animal model. The normal group and the sensorineural hearing loss (SNHL) group and the experimental (transplanted MSCs) group were compared and estimated hearing level using auditory brainstem response (ABR) recordings and the otoacoustic emission (OAE) test.

Results

ABR threshold value and DPOAE level showed that MSCs transplantation groups was improved than the SNHL group. And the number of spiral ganglion neurons were increased in all turn of the cochlea. And there was no evidence of acute immunological rejection and inflammation response was not observed.

Discussion

This study is to evaluate regenerative efficacy of hearing loss by transplanting mesenchymal stromal cells (MSCs) derived from human placenta (amnion and chorion) in deaf animal model. We identified that MSCs transplantation restored auditory impairment and promoted cell regeneration. We hope to overcome sensorineural hearing loss by transplanting stem cells such as mesenchymal stromal cells (MSCs) from easily accessible adult stem cell source in placenta.

Introduction

There are over 250 million patients worldwide suffering from incurable neurological disease, including deafness. Among patients suffering from deafness, almost 90% have some form of sensorineural dysfunction, such as sensory hair cell loss. It is estimated that the number of patients with neuron-related disease will double by the year 2030. Recently, the transplantation of stem cells has emerged as a promising therapeutic strategy for the treatment of permanent hearing loss [1], [2]. Furthermore, articles and broadcasts have reported that many incurable neurological diseases have been influenced by the use of in vivo therapeutic methods.

Mesenchymal stromal cells (MSCs) are stem cells with a capacity for self-renewal and can differentiate into mesoderm- and non-mesoderm-derived tissues. From a therapeutic perspective, MSCs are promising therapeutic agents for tissue regeneration due to their ease of preparation and immunologic privilege. Many other studies have isolated adult mesenchymal stromal cells from bone marrow, adipose tissue, umbilical cord blood (UCB), lung tissue, and Wharton's jelly [3], [4], [5], [6], [7]. Additionally, in our previous study, we demonstrated that UCB-derived mesenchymal stromal cells can differentiate into auditory hair cells and neuronal cells in vitro when exposed to a neuronal-developmental microenvironment. These cells can be differentiated into all three germ cell layers (neuron, supporting cells, and hair cells). Several studies have shown that mesenchymal stromal cells can be collected from placenta and umbilical cord blood as a derivative and used in the field of regenerative medicine [7], [8]. However, the amount of MSCs derived from UCB was very small, and it was difficult to achieve a proper environment for the differentiation of MSCs; consequently, they did not proliferate well. Recently, the isolation of MSCs from adult stromal cells has demonstrated that these cells are a potential source of mesenchymal stromal cells with therapeutic potential. Also, our previous studies and other articles have shown that many types of adult stem cells can differentiate into various cell types. In addition, these cells have been used in therapeutic pathways that promote wound-healing and effect incurable diseases and nerve disorders [7], [8], [9], [10], [11], [12], [13]. Consequently, the aim of this article was to evaluate the therapeutic regeneration and treatment effects with respect to hearing loss using mesenchymal stromal cells derived from human placenta. Also, it is mandatory to develop novel techniques for isolating, culturing, and mass producing MSCs with proliferative capacity.

Therefore, in this study, we have shown that human amniotic membrane-derived MSCs (AM-MSC) and chorionic membrane-derived MSCs (CM-MSC) can be differentiated into neurons, glial cells, and hair cells. In addition, we transplanted AM-MSC into an animal model of deafness and evaluated their therapeutic efficacy with respect to hearing.

Section snippets

Isolation of MSCs derived from human placenta

Placentas were collected after birth from women who carried to full-term after obtaining informed consent and were placed in heparin for storage. The average age of the pregnant women whose placentas were used in this study was 30–40 years. Placentas were isolated from a total of 13 women. These samples were washed three times with PBS and stored in serum-free α-MEM (Alpha Minimum Essential Medium, Gibco, Grand Island, NY). Samples were carefully divided into amnion membrane and chorionic

Isolation and differentiation of MSCs

After primary culture for 5 days, the culture dish contained mononuclear cells attached to the plate surface and in suspension. As the numbers of sub-cultures increased, the shapes of cells elongated and changed into those of fibroblast-like cells (Fig. 1A and D). All MSCs derived from amnion and chorion were maintained for at least 10 passages. Also, after each of the MSC samples was cultured in neurosphere differentiation medium containing bFGF and EGF, many sphere-forming floating cells were

Discussion

The placenta is an extra-embryonic organ with a rich source of progenitor or stem cells. The placenta has two sides; one is the fetal side consisting of amnion and chorion, and the other is the maternal side consisting of decidua [15], [16]. In this study, we demonstrated the proliferation, differentiation, and cellular restoration capacities of MSCs using amnion- and chorion-derived MSCs. MSCs isolated from the maternal side of human placenta represent an important cell type for stem cell

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    Kicheol Kil & Mi Young Choi were co-first author & equally contributed to this article.

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