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Experimental study of type Ⅰ collagen on rabbit bone marrow mesenchymal stem cell osteogenic differentiation

Author: ZhangWenPeng
Tutor: YeFaGang
School: Qingdao University
Course: Surgery
Keywords: mesenchymal stem cells cell culture collagen typeⅠ osteogenic differentiation rabbit
CLC: R329.2
Type: Master's thesis
Year: 2009
Downloads: 117
Quote: 0
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Abstract


Objective To further explore an effective way to separate and purify the rabbit bone marrow mesenchymal stem cells(rBMSCs)through experiments in vitro,and to observe the biological characteristics of the cultured cells,and to observe the role of the typeⅠcollagen protein in process of the rBMSCs osteogenic differentiation.Methods Our experiment was achieved in the Orthopedic Institute of the Affiliated Hospital of Medical College of Qingdao University in March to July 2008.rBMSCs were separated by density gradient centrifugation;Cells were cultured,passaged,purified and amplified by the adherent culture method and observed the biological characteristics;the growth of rBMSCs was observed under an inverted microscope.The ultrastructure of rBMSCs was observed by transmission electron microscope,rBMSCs surface markers were identified with the flow cytometry.After the rBMSCs were cultured on the purified typeⅠcollagen protein,the expression of the alkaline phosphatase(ALP) and ability to form mineralized nodus were examined.Results After cultured for 1 day the separated rBMSCs began to adhere,the adherent rBMSCs began to form clone at about average 10 days.The cells obtained homogeneous cell morphology,were fibroblast-like growth and stably expressed rBMSCs surface markers.After the rBMSCs were cultured on the typeⅠcollagen protein surface in vitro, the alkaline phosphatase expression was increased obviously.Mineralized nodus appeared and Von.Kossa staining presented strong positive.Conclusion To separate rBMSCs with density gradient centrifugation and to culture the rBMSCs with adherent culture method is effective in the separation and purification of rBMSCs.Cells obtained with this method have general characteristics of rBMSCs.TypeⅠcollagen protein can promote the osteogenic differentiation of rBMSCs.

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