原代培养大鼠前列腺细胞建立前列腺增生筛药模型
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国家自然科学基金项目(面上项目,重点项目,重大项目);国家科技攻关计划


Establishment of BPH drug screening model with primary cultured prostatic cells from rats in vitro
Fund Project:

The National Natural Science Foundation of China (General Program, Key Program, Major Research Plan);The National Key Technologies R&D Program of China

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    摘要:

    【摘要】目的:建立原代培养大鼠前列腺上皮细胞体外筛药模型。方法:无菌状态下取雄性SD大鼠腹侧叶前列腺,称量后,剪成1mm3小块,经Ⅱ型胶原酶消化1h后,过滤、离心获取前列腺细胞,接种于24孔板培养并对细胞进行形态学和免疫组织化学鉴定。获取的大鼠前列腺上皮细胞分别接种于96孔板和24孔板培养12d后给予系列剂量(0.1-1000µM)的他莫昔芬和阳性药物爱普列特处理72 h,利用CCK-8法检测前列腺上皮细胞存活率并计算药物IC50值,并进一步通过Giemsa染色后观察细胞生长及形态变化。结果:细胞鉴定结果显示,获取的细胞具典型上皮细胞形态特征,细胞角蛋白和前列腺特异性抗原表达阳性,提示所获细胞为前列腺上皮细胞。爱普列特与前列腺上皮细胞孵育72h后,CCK-8法检测结果显示能够明显抑制前列腺上皮细胞生长,其IC50值为42.7µM,进一步镜下观察结果显示,爱普列特未明显改变大鼠前列腺上皮细胞形态,但能导致存活细胞数减少。他莫昔芬则对大鼠前列腺上皮细胞生长无明显抑制作用,镜下观察前列腺上皮细胞数量和形态未见明显改变。结论:利用原代培养SD大鼠前列腺上皮细胞可以成功建立前列腺增生体外筛选模型。

    Abstract:

    [Abstract] Objective To establish drug screening model with primary cultured prostatic epithelial cells from rats in vitro. Methods The ventral prostate was freed from adult SD rat under sterile conditions and weighted, then cut into 1mm3 slices, after being digested with collagenase 2 for 1 h, the prostate cells were harvested through filtering and centrifuging. All cells were seeded into 24-well plate and cultured for cell morphology and immunohistochemistry detection. The prostatic epithelial cells were seeded respectively into 96-well plate and 24-well plate, treated with epristeride (0.1-1000µM)and tamoxifen (0.1-1000µM)for 72 h after being cultured for 12 d. The prostatic epithelial cell survival rate was detected by CCK-8 method, and the IC50 value of epristeride and tamoxifen was calculated, then further observed their effects on the growth and cell morphology of prostatic epithelial cells. Results According to cell morphology and immunohistochemical detection result, all cells from rat prostate displayed typical epithelial cell shape, and their expressions of cytokeratin antibody and prostate specific antigen were positive, such results suggested that the harvested cells were prostatic epithelial cell. After the prostatic epithelial cell being treated with epristeride for 72 h, its growth significantly been inhibited, and the IC50 value of epristeride was 42.7µM. Cell morphology results further showed that epristeride didn’t significantly changed the shape of prostatic epithelial cell, but it could reduce the survival prostatic epithelial cells number. While tamoxifen didn’t inhibit the cell growth, and had no obvious effect on cell number and cell shape. Conclusion Benign prostatic hyperplasia drug screening model could be successfully establish with primary cultured prostatic epithelial cells from rats in vitro.

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吴建辉.原代培养大鼠前列腺细胞建立前列腺增生筛药模型[J].中国实验动物学报,2013,21(5).

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  • 收稿日期:2013-04-24
  • 最后修改日期:2013-06-19
  • 录用日期:2013-06-24
  • 在线发布日期: 2013-10-18
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