Osteogenic Lineage Commitment of Adipose-Derived Stem Cells Is Predetermined by Three-Dimensional Cell Accumulation on Micropatterned Surface

Abstract

Lineage commitment of stem cells is mainly regulated by their microenvironments, which comprise soluble growth factors, extracellular matrix, mechanical forces, and cell density. Although numerous studies have investigated stem cell response to these factors in two-dimensional (2D) culture, little is known about that in 3D culture. Here, we studied effects of 3D cell accumulation levels on the differentiation behavior of mesenchymal stem cells (MSCs) by using a micropatterned surface. After induction of 3D-cultured MSCs on the surface, their osteogenic differentiation was significantly promoted, while adipogenic differentiation was not. This differentiation behavior of densely packed MSCs in 3D culture is unlike that in 2D culture. Moreover, to determine the contributing factor of this commitment, the relationship between 3D cell accumulation levels and their differentiation potential was studied before differentiation induction. A series of MSCs with varied 3D accumulation levels were constructed on the micropatterned surface, where the accumulated MSCs were not in hypoxic environment. Interestingly, with increasing 3D accumulation levels, MSCs enhanced their osteogenic potential but repressed adipogenic potential in the gene expression level. These results suggest that preconditioned 3D microenvironments with high cell accumulation levels promote osteogenic differentiation of MSCs and their accumulation levels help in regulating MSC differentiation.

Donor Site Location Is Critical for Proliferation, Stem Cell Capacity, and Osteogenic Differentiation of Adipose Mesenchymal Stem/Stromal Cells: Implications for Bone Tissue Engineering

“…The supplementation of the culture medium with platelet lysate or platelet-rich plasma also improved the osteogenic differentiation of Ad-MSCs [ 14 , 22 , 23 ]. Also, the cultivation on three-dimensional scaffolds further improved the osteogenic properties of Ad-MSCs [ 24 , 25 ], amongst others, through the control of cell agglomeration [ 26 ]. Furthermore, co-cultivation with endothelial cells, vascular cells, or osteoblastic cells improved the osteogenic differentiation of Ad-MSCs [ 27 , 28 , 29 ].…”

Chemistry Can Make Strict and Fuzzy Controls for Bio-Systems: DNA Nanoarchitectonics and Cell-Macromolecular Nanoarchitectonics

“…In the case of MSC, effective reduction of differentiation induction time for osteoblast cells was accomplished using microdomain patterned surface, where cell accumulations accelerate the osteogenic differentiation in gene expression level (Figure 31). 444 These results indicate the importance of three-dimensional cell architectures constructed on the two dimensional microdomain patterned surfaces.…”

Airflow‐Assisted 3D Bioprinting of Human Heterogeneous Microspheroidal Organoids with Microfluidic Nozzle

“…10 569 096) with 10% FBS, 50 µg mL −1 ascorbate-2-phosphate, 10-8m dexamethasone and 10 × 10 −3 m β-glycerophosphate. [39][40][41][42] To enhance the angiogenesis of HUVECs, 10 ng mL −1 human VEGF (Peprotech, Cat. #100-20A-10UG) was added in the medium.…”

“…Osteogenic Induction and Characterization : Cell‐laden spheroids were cultured in osteogenic differentiation medium consisted of H‐DMEM (Gibico, Cat. 10 569 096) with 10% FBS, 50 µg mL −1 ascorbate‐2‐phosphate, 10–8 m dexamethasone and 10 × 10 −3 m β‐glycerophosphate . To enhance the angiogenesis of HUVECs, 10 ng mL −1 human VEGF (Peprotech, Cat.…”