雌激素对鸡胚BMSCs诱导的成骨细胞增殖、凋亡及细胞周期的影响

doi:10.11843/j.issn.0366-6964.2014.02.005

Abstract

Abstract:

To investigate the effect of 17β-E₂on osteoblasts induced by chicken bone marrow mesenchymal stem cells(BMSCs) ，the method of in vitro culture of BMSCs and its inducement of osteoblasts were established in this experiment.Chicken BMSCs were purified with whole marrow adherent culture and subculture in vitro.The third generation of cells of BMSCs were selected for inducing osteoblastic culture.The growth and morphology，the cell proliferation rate and the cell cycle of osteoblasts induced-BMSCs were detected with inverted phase contrast microscope，methyl-thiazol-tetrazolium (MTT) and flow cytometry，respectively.Induced-BMSCs obsteoblasts were identified by alkaline phosphates (ALP)，Van-Gieson staining and Alizarin red staining，respectively.The induced osteoblasts were divided into 5 experimental groups and cultured with osteogenic differentiated medium containing different 17β-E₂ concentrations of 10^-7，10^-8，10^-9，10^-10，10^-11 mol·L^-1 for each group，respectively.Cells cultured without 17β-E₂ as a blank control group.Effects of different concentrations of 17β-E₂on the proliferation and differentiation of induced-BMSCs osteoblasts were measured by MTT and PNPP for finding the most effective concentration.Then this effective concentration of 17β-E₂was used for culturing induced-BMSCs osteoblasts，and the percentages of apoptotic cells were measured by flow cytometry.The results showed that BMSCs could be purified with whole marrow adherent culture.Osteoblasts could be induced from purified BMSCs using classic differentiated medium and were typically characteristic of the osteoblastic biology.The induced-BMSCs osteoblasts’ ALP，collagen typeⅠand calcified nodules’ staining were strongly positive compared with control group.10^-9 mol·L^-1 17β-E₂concentration represented the most action of promoting proliferation and differentiation and inhibiting apoptosis of induced-BMSCs osteoblasts among the other concentration groups.Thus，it is shown that osteoblasts can be induced from purified chicken BMSCs and estrogen can enhance the function of induced-BMSCs osteobalsts.

CLC Number:

S831
S814.1

YU Ze-zhong，WANG Zhi-mei，JIANG Sha，HOU Jia-fa. Effect of 17β-E₂ on Proliferation，Apoptosis and Cell Cycle of Osteoblasts Induced by BMSCs in Chicken Embryos[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(2): 197-206.

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Effect of 17β-E₂ on Proliferation，Apoptosis and Cell Cycle of Osteoblasts Induced by BMSCs in Chicken Embryos

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Effect of 17β-E2 on Proliferation，Apoptosis and Cell Cycle of Osteoblasts Induced by BMSCs in Chicken Embryos

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Effect of 17β-E₂ on Proliferation，Apoptosis and Cell Cycle of Osteoblasts Induced by BMSCs in Chicken Embryos