The Effect Of Fluid Shear Stress (FSS) On Human Mesenchymal Stem Cells Proliferation And Osteogenic Differentiation

Bioreactor is very hot in the area of bioengineering; it is widely used in culturing cells and tissue. Perfusion bioareactor has advantages in bone tissue engineering, because of the dynamic 3D culture pattern. The dynamic environment can provide fluid shear stress(FSS) on the cultured cells. In order to maintain the mass and the normal functions of the bone, it needs to put suitable stress on the bone. When the interstitial fluid flow was driven by the formation of the bone, it will provide FSS on the cells of the bone. More evidences has supported the fact that FSS is the main stress on the bone cell. And FSS has very important effect on the proliferation and differentiation.Parallel-plate flow chamber(PPFC) is a experimental facility that can provided FSS of precise magnitude, so it is widely used to study the hydrodynamics of cell. Recently, mensenchymal stem cells(MSCs) is considered one of the most developmental seed cells of bone tissue engineering. Human mesenchymal stem cells is established on the matrix of bone marrow, and the environment is very familiar with bone cells. So we presume that hMSCs is also sensitive to FSS, and the familiar effect can be made when take FSS on hMSCs. In our work, we upgrade the old type of PPFC, and stimulate the hMSCs with FSS using the upgraded PPFC. We observe the form change of hMSCs through inverted microscope and TEM; through MTT and flow cytometer to observe the proliferation; through the ALP Staining, Calcium staining for certain dyestuff, Tetracycline fluorescence labelling, estimating the excretion of osteocalcin by radioimmunoassay and the activity of the cellular ALP to observe the differentiation.The objective we do the study is to observe the effect of FSS on the proliferation and differentiation of hMSCs. And it can establish the foundation to study the mechanism of the FSS on hMSCs in the future. It also can provide theory and experiment basic for the use of perfusion bioreactor on culturing tissue engineering bone. From the experiment, we get the following result:...