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To Discuss the Mechanism and the Function of Nucleostemin by Differentiating Rat Bone Marrow stem Cells into Osteoblasts in Vitro

Author: PeiFang
Tutor: WangJianChao
School: Huazhong University of Science and Technology
Course: Orthopedics
Keywords: Bone marrow mesenchymal stem cells Induced differentiation Osteoblasts Nuclear stem cell factor
CLC: R329
Type: Master's thesis
Year: 2010
Downloads: 35
Quote: 0
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Abstract


Objective: To investigate Nucleostemin (NS) in rat bone marrow mesenchymal stem cells induced differentiation mechanism and the role of osteoblasts . The method : Adapted chosen 4 to 6 -week-old male SD rats bilateral tibial and femoral bone marrow stromal cells , and primary and subculture . Cell culture to the 3rd generation of a control group , were mixed bone marrow mesenchymal stem cells cultured in medium with normal culture medium and mineralization . Turn by phase contrast inverted microscope cell growth , MTT assay of cell proliferation , alkaline phosphatase (AKP) and calcium alizarin red staining to analyze the activity of the cells into bone . Immunohistochemical SABC method and immunofluorescence assay NS cells , respectively expression in normal medium and osteoblast induction medium under comparison NS . Results: mineralization medium induced rat bone marrow mesenchymal stem cells compared with ordinary culture medium of expression of NS expression was significantly weakened, and in a time -dependent attenuation . Bone marrow mesenchymal stem cells can be induced in the role of the mineralized solution for slug distribution osteoblasts . Osteoblasts NS expression was negative . MTT assay demonstrated that mineralized solution to develop a long incubation period, cell growth , compared with the control group to extend the logarithmic growth phase . Conclusion: ① This method can get a lot of mesenchymal stem cells in the bone marrow . ② bone marrow mesenchymal stem cells differentiate into osteoblasts through mineralization induced fluid . The ③ NS as a nuclear gene regulatory fragment of stem cell factor , protein table to the amount is quite rich in stem cells . , And its expression is significantly reduced when the cells differentiate . ④ Therefore , NS genes directly involved in the regulation of cell proliferation and is likely to serve as a symbol of rat bone marrow mesenchymal stem cells have the potential ability to differentiate factor .

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