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

Abstract

Background: Adipose-derived mesenchymal stem cells (ADMSCs) have attracted widespread interest as cell-based tissue repair systems. To obtain adequate quantities of ADMSCs for therapeutic applications, extensive in vitro expansion is required. However, under current two-dimensional (2D) approaches, ADMSCs rapidly undergo replicative senescence, and cell growth is impeded and stem cell properties are eliminated by mechanisms that are poorly understood. These issues limit the extensive applications of ADMSCs. In this study, we investigated senescence-related changes in mesenchymal stem cells (MSCs) isolated from human adipose tissue in 2D and three-dimensional (3D) cultures. Methods: We studied cell growth over a given period (21 days) to determine if modes of culture were associated with ADMSC senescence. ADMSCs were isolated from healthy females by liposuction surgery and then were grown in 2D and 3D cultures. The cell morphology was observed during cell culture. Every other time of culture, senescence-associated β-galactosidase (SA-β-gal) expression, cell viability, proliferation, and differentiation potential of ADMSCs from 2D and 3D cultures were detected. Also, senescence-and stemness-related gene expression, telomere length, telomerase activity, and energy metabolism of ADMSCs for different culture times were evaluated. Results: With long-term propagation, we observed significant changes in cell morphology, proliferation, differentiation abilities, and energy metabolism, which were associated with increases in SA-β-gal activity and decreases in telomere length and telomerase activity. Notably, when cultured in 3D, these changes were improved. Conclusions: Our results indicate that 3D culture is able to ameliorate senescence-related changes in ADMSCs.

Impact of 3D cell culture on bone regeneration potential of mesenchymal stromal cells

“…In contrast, a follow-up study showed that an anionic form of nanofibrillar cellulose significantly increased the viability of human AD-MSCs [ 71 ]. Similarly, Yin and colleagues recently reported that 3D cultivation of AD-MSCs in a nanofibrous polysaccharide hydrogel increases their proliferation and viability [ 88 ].…”

“…Moreover, an overall higher telomerase activity and greater telomere length was observed in MSCs in 3D. This was accompanied by a higher adipogenic and osteogenic differentiation potential [ 88 ].…”

Electrical Stimulation of Adipose-Derived Stem Cells in 3D Nanofibrillar Cellulose Increases Their Osteogenic Potential

“…In this context, it has been reported that 2D culture can result in a loss of multipotency, premature cellular senescence, and accumulation of chromosomal aberrations within the MSC genome [31][32][33]. In contrast, 3D cell culture not only mimics the 3D structure of the bone tissue but can also positively influence osteogenic differentiation of MSCs whilst simultaneously reducing their cellular senescence [34]. MSCs of different tissue origin have been successfully cultivated in a multitude of different solid 3D scaffolds and hydrogels including calcium phosphate ceramic [35], apatite-wollastonite [36], poly(ε-caprolactone) [37], alginate [38] collagen [39], Matrigel [40], bacteria-derived cellulose [41], and methylcellulose [42].…”

Assessment of Long-Term in vitro Multiplied Human Wharton’s Jelly-Derived Mesenchymal Stem Cells prior to Their Use in Clinical Administration

“…The relative telomere length was found to be decreased with increase of sequential passage of WJ-MSCs. Former studies by Samsonraj et al [2013], Kim et al [2017], and Yin et al [2020] have also observed gradual decrease in telomere length of BM-MSCs, PBMSCs and AD-MSCs, respectively.…”