Visualization of Src and FAK activity during the differentiation process from HMSCs to osteoblasts.
Non-receptor protein kinases FAK and Src play crucial roles in regulating cellular adhesions, growth, migration and differentiation. However, it remains unclear how the activity of FAK and Src is regulated during the differentiation process from mesenchymal stem cells (MSCs) to bone cells. In this study, we used genetically encoded FAK and Src biosensors based on fluorescence resonance energy transfer (FRET) to monitor the FAK and Src activity in live cells during the differentiation process. The results revealed that the FAK activity increased after the induction of differentiation, which peaked around 20-27 days after induction. Meanwhile, the Src activity decreased continuously for 27 days after induction. Therefore, the results showed significant and differential changes of FAK and Src activity upon induction. This opposite trend between FAK and Src activation suggests novel and un-coupled Src/FAK functions during the osteoblastic differentiation process. These results should provide important information for the biochemical signals during the differentiation process of stem cells toward bone cells, which will advance our understanding of bone repair and tissue engineering.
Biomaterials and Live Cell Imaging Institute, School of Metallurgy and Materials Engineering, Chongqing University of Science and technology, Chongqing, People's Republic of China.
Xiaoling
X
Liao
Shaoying
S
Lu
Yue
Y
Zhuo
Christina
C
Winter
Wenfeng
W
Xu
Yingxiao
Y
Wang
D000469:Alkaline Phosphatase / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D004789:Enzyme Activation; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology; D010006:Osteoblasts / Q000166:cytology* / Q000201:enzymology; D010012:Osteogenesis; D013056:Spectrophotometry, Ultraviolet; D019061:src-Family Kinases / Q000378:metabolism*
D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases; D000469:Alkaline Phosphatase