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The Experimental Study of the Proliferation and Osteogenic Differention Effects of Zuogui Pill on Rat Mesenchymal Stem Cells

Author: YanChunLu
Tutor: WangShuanKe
School: Lanzhou University
Course: Surgery
Keywords: The Zuogui Pill Containing serum Bone marrow mesenchymal stem cells Osteoblasts
CLC: R285.5
Type: Master's thesis
Year: 2008
Downloads: 77
Quote: 0
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Abstract


Objective: observed Zuo Gui Pill on cultured rat bone marrow mesenchymal stem cells (Mesenchymal StemCells, MSCs) proliferation and differentiation of osteoblasts. Method: 1. Use of whole bone marrow method (adherence screening method) isolation and culture of rat MSCs were identified; 2. Preparation of high school low concentration Zuogui Pill containing serum was detected by MTT the Zuogui Pill MSCs proliferation square containing serum understanding of the dose-effect relationship; Zuo Gui Pill with different concentrations of the drug-containing serum induce MSCs to differentiate into osteoblasts (osteoblast, OB), the points high concentration of Zuoguiwan plus Less party group, Zuo Gui Pill group concentration, low concentration Zuo Gui Pill group, classic induction group and blank control group of five groups; Von-Kossa staining of different concentrations of Zuoguiwan plus Less square containing serum induced cells were stained determine the Pill Zuogui containing serum induced osteogenesis whether MSCs; 5. using immunocytochemistry to detect the expression of type I collagen, and gray-scale analysis Pill determine Zuogui containing serum on whether the MSCs osteogenic differentiation-inducing effect. Results: 1.MSCs separated basic adherent, 12 ~ 15d achieve fusion; 2. Flow cytometry results showed that, in the third generation of MSCs G 0 / G 1 96.7% of the cells, G 2 of the cells were 1.2%, 2.1% in the S phase of the cell; low expression of CD29, high expression of CD44 and CD105, no expression of CD34, CD45, CD19, in line with the characteristics of the mesenchymal stem cells in the bone marrow; 3. compared with the blank control group, different concentrations of Zuogui Pill containing serum can promote the proliferation of MSCs, and its effect was dose dependent (P <0.05); by Von-Kossa staining can join the different concentrations Zuogui Pill containing serum induced 21d observed black mineralized nodules; 5. compared with the blank control group, in addition to the low concentration Zuogui Pill side drug-containing serum-induced the 7d on of collagen Ⅰ average gray expression not significantly affect (P> 0.05), different concentrations Zuogui Pill containing serum upregulate the expression of the of collagen Ⅰ average gray are (P <0.05). Conclusion: low high school concentration Zuogui of Pill containing serum has a role to promote the proliferation of MSCs, and was dose dependent. Each concentration Zuo Gui Pill containing serum could significantly induce their differentiation into osteoblasts (OB), so Zuo Gui Pill to prevent and treat osteoporosis, and possible mechanisms of proliferation by promoting MSCs or direct induction differentiation into osteoblasts (OB), thereby enhancing bone of MSCs differentiation capacity.

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