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Experimental Study of Induction of Mouse Bone Marrow Derived Stem Cells into Osteoblast Like Cells with Periosteum Extract

Author: ZhangLiLi
Tutor: WangZhiYing
School: Liaoning Medical
Course: Clinical Stomatology
Keywords: Bone marrow mesenchymal stem cells Periosteum extract Induced differentiation Osteoblasts
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 32
Quote: 0
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Abstract


Purpose to induce bone marrow mesenchymal stem cells (BMSCs) the differentiation site-specific , application mouse periosteum extract of mouse bone marrow mesenchymal stem cell differentiation observed induced differentiation . Method to the skull periosteum extract in mice and mouse bone marrow -derived mesenchymal stem cells in vitro co-culture -induced murine bone marrow -derived mesenchymal stem cells to the class of osteoblast differentiation . The experiment was divided into the control group (only BMSCs ) and experimental group ( the periosteum extract induced differentiated cells ) , inverted microscope induced changes in cell morphology ; application of alkaline phosphatase ( ALP ) staining induced to differentiate into cells the bone activity ; application immunoblotting BMP-2 protein expression ; RT-PCR was used to detect the expression of BMP - 2 , type I collagen , osteocalcin gene . Results of morphological observation , BMSC cells fusiform periosteal induced to differentiate into a triangle , polygon , cubic ; ALP activity after induction of differentiation ALP activity is much higher than BMSC cells , which is about six times its both The difference was statistically significant ( P lt ; 0.01 ) ; Western blot results showed that the group of BMSC cells do not express BMP-2 protein , periosteum extract induced differentiation of BMP-2 protein expression increased , the difference was statistically significant ( P lt ; 0.01 ) ; RT - PCR results showed that mice BMSC cells do not express BMP - 2 , type I collagen , osteocalcin gene . Periosteum extract induced differentiation can be detected BMP - 2 ( 708bp ) , type I collagen ( 222bp ) , osteocalcin ( 219 bp ) gene expression . beta - actin as internal reference . Conclusion periosteum extract can induce murine bone marrow -derived mesenchymal stem cell differentiation of osteoblast cells , differentiated cells with osteoblast characteristics of this method is simple and works well differentiated for bone tissue engineering to provide a new class osteoblast culture method .

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