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 ADMSCs senescence . ADMSCs were isolated from healthy females by liposuction surgery and then were grew 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 genes expression, telomere length, telomerase activity, and energy metabolism of ADMSCs for different culture time 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

“…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].…”

“…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].…”