Abstract

Mesenchymal stem cells (MSC) exhibit enormous heterogeneity which can modify their regenerative properties and therefore influence therapeutic effectiveness as well as safety of these cells transplantation. In addition the high phenotypic plasticity of MSC population makes it enormously sensitive to any changes in environmental properties including fluctuation in oxygen concentration. We have shown here that lowering oxygen level far below air atmosphere has a beneficial impact on various parameters characteristic for umbilical cord Wharton Jelly- (WJ-) MSC and adipose tissue- (AD-) derived MSC cultures. This includes their cellular composition, rate of proliferation, and maintenance of stemness properties together with commitment to cell differentiation toward mesodermal and neural lineages. In addition, the culture genomic stability increased significantly during long-term cell passaging and eventually protected cells against spontaneous transformation. Also by comparing of two routinely used methods of MSCs isolation (mechanical versus enzymatic) we have found substantial divergence arising between cell culture properties increasing along the time of cultivation in vitro. Thus, in this paper we highlight the urgent necessity to develop the more sensitive and selective methods for prediction and control cells fate and functioning during the time of growth in vitro.

Highlights

  • Mesenchymal stem cells (MSC) cultures exhibit enormous heterogeneity which can influence their therapeutic efficacy as well as safety after transplantation

  • Stem Cells International long cell culture like failure of proliferation or cell genome transformation. This can be achieved, as we present it for adipose tissue- (AD-)MSC culture experiments, by changing oxygen environment from 21% to ≤5% O2 concentration which physiologically occurs in the majority of tissues and stem cell niches in vivo [4]

  • Wharton Jelly- (WJ-)MSC were isolated by either mechanical- (M-) or enzymatic- (E-) based procedures as described in Section 2 and cultured for 14 days under 21% O2 or 5% O2

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Summary

Introduction

MSC cultures exhibit enormous heterogeneity which can influence their therapeutic efficacy as well as safety after transplantation. Since we approached era of increasing application of MSC therapy in a clinic, standardization of cell manufacturing with optimized isolation efficacy, rate of cell proliferation, and viability and longevity of culture with capacity to differentiation into desired lineages seems to be of key practical issues. All these important demands are not adequately considered and verified by routinely used standard MSC culture preimplantation screening still based on the ISSCR criteria established by International Society for Stem Cell Research in 2008. At the end we will propose the simple and easy method to prevent appearance of the most adverse events faced during

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