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The Study about OGP and the Mechanism of OGP to Promote Rat Bone Marrow Mesenchymal Stem Cells to Osteogenetic Differentiation

Author: WangJianGuo
Tutor: BaiZengLiang
School: Shandong University
Course: Developmental Biology
Keywords: Osteogenic growth peptide Bone marrow mesenchymal stem cells Osteogenic differentiation Gene expression profiling Immunoassay method
CLC: R329
Type: Master's thesis
Year: 2008
Downloads: 176
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Abstract

Osteogenic growth peptide (Osteogenic Growth Peptide, OGP) histone H4 gene encoding a polypeptide. , OGP has extensive osteogenic and hematopoietic activity. Body OGP can stimulate bone formation and increased trabecular bone density, promote the healing of fractures; vitro the OGP can adjust variety of osteoblasts, the proliferation of bone marrow mesenchymal stem cells to improve the alkaline phosphatase activity, promote matrix mineralization. OGP can also stimulate the generation of hematopoietic cells, improve the survival rate of the bone marrow transplant, and the OGP hematopoietic activity OGP improve the hematopoietic microenvironment related. Speculated OGP osteogenic and hematopoietic activity may be related to bone marrow mesenchymal stem cells. Bone marrow mesenchymal stem cells (BMSCs) is a bone marrow have multi-differentiation potential of stem cells, is the bone marrow hematopoietic micro-environment of the important composition part, under appropriate conditions can be differentiated as osteoblasts, chondrocytes, fat cells and other. Of mesenchymal stem cells in the bone marrow hematopoietic cell growth, differentiation essential micro-environment, but also the precursor cells into bone progenitor cells, bone marrow hematopoietic and osteogenic has a very important role. In order to study the relationship of OGP the biological activity of bone marrow mesenchymal stem cells and possible application prospects of this thesis was studied from the following aspects: (1) rat bone marrow mesenchymal stem cells (rBMSCs) Isolation, culture and OGP of rat bone marrow mesenchymal stem cell proliferation and osteogenic differentiation of role. The results show that, OGP at low concentrations (10 ^{-10 mol / L, 10 ^{-11 mol / L) when the original activity of rat bone marrow mesenchymal stem cells in mitosis, high concentration (10 ^{-8 mol / L, 10 ^{-9 mol / L) to promote osteogenic differentiation of rat bone marrow mesenchymal stem cells. By RT-PCR and gene chip results show, OGP osteogenic differentiation of early promotion of rat bone marrow mesenchymal stem cells may be related to the osteoblast-specific transcription factor Cbfα1; OGP induced rat bone marrow mesenchymal stem cell osteogenic differentiation of early specific expression of osteoblast markers osteocalcin (Osteocalcin) negative expression showed that rat bone marrow mesenchymal stem cells is not yet complete differentiation to osteoblasts. (2) osteogenic growth peptide induced rat bone marrow mesenchymal stem cell gene expression changes. By Boao 27K Rat Genome Array chip, the changes the OGP induced early gene expression profiling of rat bone marrow mesenchymal stem cells. Found a total of 145 differentially expressed genes, 91 genes upregulated and 54 down-regulated expression of genes. Possible genes associated with osteogenic and hematopoietic activity differentially expressed genes in a total of 35: including four of angiogenesis-related genes, nine bone metabolism-related genes, 14 genes related to inflammation and immune cell proliferation differentiation associated gene. DNA microarray results promoting osteogenesis of rat bone marrow mesenchymal stem cells and hematopoietic activity of molecular mechanisms to establish a good foundation for further study OGP. (3) recombinant vector transfected rat bone marrow mesenchymal stem cells and their activity comparison into bone differentiation activity and osteogenic growth peptide induced rat bone marrow mesenchymal stem cells osteogenic differentiation. PEGFP-N1 plasmid, constructed of the OGP justice carriers and Pre-OGP antisense expression vector, and transfection of rat bone marrow mesenchymal stem cells. The fluorescence observation showed that the recombinant vector can be successfully transfected with rat bone marrow mesenchymal stem cells. Alkaline phosphatase (ALP) activity assay showed that antisense carrier of the Pre-OGP transfected rat bone marrow mesenchymal stem cell osteogenic differentiation activity was inhibited, OGP justice vector-transfected rat bone marrow mesenchymal stem cells are easier to osteogenic differentiation. (4) osteogenic growth peptide establishment of immunological detection methods. Delayed fracture healing and fracture healing in order to study the OGP with osteoporosis and other metabolic bone diseases, such as the relationship between clinical disease, the paper established an immunological detection methods OGP. Established a method for the clinical detection of the concentration of serum OGP through improved immunological method, compared with traditional methods, the sensitivity is greatly increased.}}}}

The Study about OGP and the Mechanism of OGP to Promote Rat Bone Marrow Mesenchymal Stem Cells to Osteogenetic Differentiation

Abstract

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