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作 者:于永忠[1] 王欢[1] 谭强[1] 赵文博[1] 崔玉东[1]
机构地区:[1]黑龙江八一农垦大学生命科学技术学院,大庆163319
出 处:《黑龙江八一农垦大学学报》2013年第4期40-44,57,共6页journal of heilongjiang bayi agricultural university
基 金:国家自然科学基金面上项目(31172353);黑龙江省教育厅科学技术研究项目(11551322)
摘 要:研究人羊膜上皮细胞(hAEC)在不同生长因子存在时生长速度差异性及在羊口疮病毒(ORFV)作用下形态学变化特征。采用胶原酶-胰蛋白酶联合消化法分离获取hAEC并进行原代培养。分别向培养液中添加40 ng·mL-1表皮生长因子(EGF)和10 ng·mL-1的碱性成纤维细胞生长因子(bFGF),倒置显微镜观察细胞生长和增殖情况,以选择最适宜hAEC原代培养方法。体外实验用ORFV感染hAEC并记录。获得大量hAEC细胞。细胞培养时添加bFGF,hAEC生长和增殖速度显著加快。病毒感染出现明显细胞病变。胶原酶-胰蛋白酶联合消化及培养液中添加10 ng·mL-1的bFGF是hAEC较为适宜的原代培养方法;hAEC对于ORFV具有敏感性。To compare the diversity of potency of two different growth factors to the human amniotic epithelial cells (hAEC) and complete the morphological changed characteristic experiment under ORFV stimulate, hAEC were obtained by collagenase and trypsin digestion method. 40 ng'mL-1 epidermal growth factor (EGF) and 10 ng'mL-l basic fibroblast growth factor (bFGF) were added separately to the culture media, and inverted microscope was used to observe the cell growth and proliferation in order to choose the most suitable methods for hAEC in primary culture, and probe susceptibility of hAEC through ORFV infection in vitro and record. Cell culture medium added with bFGF lead to significant acceleration obtained as well in hAEC growth and proliferation rate. The primary cells cultured without growth factors were served as controls.Virus infection emerged obvious cell lesion. Collagenase , trypsin digestion method and euhure medium with 10 ng mL-1 bFGF were favourable conditions for primary culture of hAEC which had high sensitivity to ORFV.
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