Hybrid complexes of high and low molecular weight hyaluronan delay in vitro replicative senescence of mesenchymal stromal cells: a pilot study for future therapeutic application.

Nicola Alessio,Giovanni Di Bernardo,Antonietta Stellavato,Stefania Del Gaudio,Umberto Galderisi,Gianfranco Peluso,Mario De Rosa,Tiziana Squillaro,Chiara Schiraldi

doi:10.18632/aging.101493

Abstract

Mesenchymal stem cells, a subpopulation of mesenchymal stromal cells (MSCs), are present in the stroma of several tissues. MSC in vitro cultivation for clinical treatments may greatly affect MSC properties. A primary handicap is replicative senescence that impairs MSC functions. Hyaluronan (HA) is present in the extracellular matrix that composes the stem cell niche environment and is under investigation as a key factor for in vitro stem cell growth. We evaluated the effect on MSC cultivation of HA hybrid cooperative complexes (HCC) that are obtained from high (H) and low (L) weight molecules (NAHYCO™). We compared this HCC with H-HA and L-HA. We investigated the effects of these HAs on proliferation, cell cycle, apoptosis, senescence, and differentiation following the addition of the polymer solutions in the culture media at concentrations that did not drastically modify the medium viscosity. Interestingly, 0,16% HCC significantly delayed the senescence compared with the controls. This occurred without alteration of the cell cycle, cytotoxicity, or apoptosis. HCCs also promoted adipogenic and chondrogenic differentiation. Our finding could suggest a potential functional role of HCC above the updated scientific reports of its effects and pave the way to optimization of MSC cultivation for therapeutic application.

Highlights

Mesenchymal stem cells are a subpopulation of mesenchymal stromal cells (MSCs) that are present in the stroma component of several organs and tissues, mainly in bone marrow and adipose tissue [1]
Due to its abundance in the stem cell niche, we evaluated the effect on MSC cultivation of HA hybrid complexes (HCC) that are obtained from high (H) and low (L) weight molecules (NAHYCOTM technology)
At the beginning of each experiment, we evaluated that MSC cultures fulfill the ISCT (International Society for Cellular Therapy) criteria and were devoid of monocytes/macrophages [9]

Summary

Introduction

According to the International Society for Cellular Therapy (ISCT) criteria, the isolation of MSCs produces heterogeneous, non-clonal cultures of stromal cells containing stem cells with different multipotential properties, committed progenitors, and differentiated cells. MSCs present immunomodulatory properties and the capacity to sustain tissue repair and homeostasis. All these features have paved the way to several potential therapeutic applications of MSCs [2]. In vitro cultivation is a necessary procedure to obtain an elevated number of MSCs for clinical purposes. This poses fundamental issues since in vitro expansion of MSCs may greatly affect their properties. The presence of senescent cells has to be carefully evaluated in order to safeguard the therapeutic potential of any MSC batch destined for patients

Methods

Results

Conclusion