Experimental Strategies of Mesenchymal Stem Cell Propagation: Adverse Events and Potential Risk of Functional Changes

Abstract

Mesenchymal stem cells (MSCs) are attractive candidates for cell-based tissue repair approaches. Hundreds of clinical trials using MSCs have been completed and many others are still being investigated. For most therapeutic applications, MSC propagation in vitro is often required. However, ex vivo culture condition is not fully physiological and may affect biological properties of MSCs including their regenerative potential. Moreover, both cell cryopreservation and labelling procedure prior to infusion may have the negative impact on their expected effect in vivo. The incidence of MSC transformation during in vitro culture should be also taken into consideration before using cells in stem cell therapy. In our review, we focused on different aspects of MSC propagation that might influence their regenerative properties of MSC. We also discussed the influence of different factors that might abolish MSC proliferation and differentiation as well as potential impact of stem cell senescence and aging. Despite of many positive therapeutic effects of MSC therapy, one has to be conscious about potential cell changes that could appear during manufacturing of MSCs.

Comparison of senescence-related changes between three- and two-dimensional cultured adipose-derived mesenchymal stem cells

“…In terms of morphological and biochemical characteristics, in vitro MSCs senescence is characterized by an enlarged flat cellular morphology, increased SA-β-gal activity, increased expression of p16, p21 and p53 [12,35], and a loss of stem cell properties [36]. Studies have reported that MSCs enter replicative senescence after extensive culture, leading to morphological and functional cellular changes [23,37]. Cells senescence is a major factor that affects the proliferation and multi-lineage potential of MSCs [23,38].…”

“…Thus, telomere shortening more than likely functions as a mechanism implicated in cellular senescence [39,41]. Reports have identified a direct correlation between telomerase activity and stem cell function [37], with telomere attrition contributing to aging. [42] Furthermore, studies have demonstrated that the relative telomere length of ADMSCs decline with long-term passaging [38].…”

Revealing Cellular Heterogeneity andIn VitroDifferentiation Trajectory of Cultured Human Endometrial Mesenchymal-like Stem Cells Using Single-cell RNA Sequencing

“…To obtain large amount MSCs for clinical use requires in vitro expansion because of limited supply of primary MSCs. However, in vitro expansion has large inter-sample variations and causes functional loss of the cells [43][44][45][46]. Thus, understanding of in vitro differentiation is critical.…”

Comparison of senescence-related changes between three- and two-dimensional cultured adipose-derived mesenchymal stem cells

“…and p53 [12,35], and a loss of stem cell properties [36]. Studies have reported that MSCs enter replicative senescence after extensive culture, leading to morphological and functional cellular changes [23,37]. Cell senescence is a major factor that affects the proliferation and multi-lineage potential of MSCs [23,38].…”