Reference - Detail
|Author||Komatsu N, Kajiya M, Motoike S, Takewaki M, Horikoshi S, Iwata T, Ouhara K, Takeda K, Matsuda S, Fujita T, Kurihara H.|
|Title||Type I collagen deposition via osteoinduction ameliorates YAP/TAZ activity in 3D floating culture clumps of mesenchymal stem cell/extracellular matrix complexes.|
|Journal||Stem Cell Res Ther|
BACKGROUND:Three-dimensional (3D) floating culture clumps of mesenchymal stem cell (MSC)/extracellular matrix (ECM) complexes (C-MSCs) consist of cells and self-produced ECM. Previous studies have demonstrated that C-MSCs can be transplanted into bony lesions without an artificial scaffold to induce bone regeneration. Moreover, osteoinductive medium (OIM)-treated C-MSCs (OIM-C-MSCs) have shown rapid and increased new bone formation in vivo. To apply OIM-C-MSCs for novel bone regenerative cell therapy, their cellular properties at the molecular level must be elucidated. The transcriptional co-activators yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) have been recognized as key players in the mechanotransduction cascade, controlling cell lineage commitment in MSCs. It is plausible that 3D C-MSCs/OIM-C-MSCs cultured in floating conditions could provide distinct microenvironments compared to conventional 2D culture systems and thereby induce unique mechanotransduction cascades. Therefore, this study investigated the YAP/TAZ activity in 3D-cultured C-MSCs/OIM-C-MSCs in floating conditions.
METHODS:Human bone marrow-derived MSCs were cultured in growth medium supplemented with ascorbic acid. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and were then torn off. The sheet was rolled to make round clumps of cells. Then, YAP/TAZ activity, filamentous actin (F-actin) integrity, collagen type I (COL1) production, and the differentiation potency in 3D floating culture C-MSCs/OIM-C-MSCs were analyzed.
RESULTS:C-MSCs cultured in floating conditions lost their actin cytoskeleton to downregulate YAP/TAZ activity, which directed cells to undergo adipogenesis/chondrogenesis. OIM treatment induced abundant COL1 deposition, which facilitated Intβ1-dependent actin fiber formation and YAP/TAZ activity to elevate the expression levels of osteogenic master transcriptional factor runt-related transcription factor 2 (RUNX2) mRNA in C-MSCs. Importantly, elevation of YAP/TAZ activity via OIM was associated with COL1 deposition and F-actin integrity, suggesting a positive feedback loop in OIM-C-MSCs.
CONCLUSION:These findings suggest that OIM-C-MSCs, which form a unique microenvironment that maintains high YAP/TAZ activity, can serve as better candidates for bone regenerative cell therapy than C-MSCs.
|MeSH||Actin Cytoskeleton / drug effects Actin Cytoskeleton / metabolism Actins / metabolism Adaptor Proteins, Signal Transducing / metabolism* Adipogenesis / drug effects Cell Aggregation / drug effects Cell Culture Techniques / methods* Cells, Cultured Chondrogenesis / drug effects Collagen Type I / metabolism* Culture Media / pharmacology Down-Regulation / drug effects Extracellular Matrix / drug effects Extracellular Matrix / metabolism* Feedback, Physiological Humans Integrin beta1 / metabolism Intracellular Signaling Peptides and Proteins / metabolism* Mechanotransduction, Cellular Mesenchymal Stem Cells / cytology* Mesenchymal Stem Cells / drug effects Mesenchymal Stem Cells / metabolism* Models, Biological Osseointegration* / drug effects Osteogenesis / drug effects Phosphoproteins / metabolism* Signal Transduction rho-Associated Kinases / metabolism|
|Human and Animal Cells||HuCCT1(RCB1960)|