欢迎来到《四川大学学报(医学版)》
蒋 玲, 胡远东, 徐菲菲, 等.新生大鼠Ⅰ型肺泡上皮细胞原代培养方法的改进.四川大学学报(医学版),2017,48(2):282-285
新生大鼠Ⅰ型肺泡上皮细胞原代培养方法的改进
The Improvement in the Primary Culture of Alveolar Epithelial Type of Rats
  
中文关键词:  【关键词】 Ⅰ型肺泡上皮细胞 原代培养 鉴定
英文关键词:【Key words】 Alveolar epithelial typeⅠ Primary culture Identification
基金项目:
摘要点击次数: 1967
全文下载次数: 0
中文摘要:
      【摘要】 目的 改进体外SD大鼠Ⅰ型肺泡上皮细胞(alveolar epithelial typeⅠ,ATⅠ)的原代培养方法,用更为简单的培养方法获得较高纯度的ATⅠ。方法 选用新生1d的SD大鼠,消毒后待新生大鼠呼吸停止后断头取肺,采用胶原酶Ⅰ和DNaseⅠ消化分离法获取原代细胞,将细胞接种在Ⅰ型鼠尾胶原蛋白包被过的细胞板中,利用差速贴壁法纯化AT Ⅰ,同时细胞培养48 h后进行换液,倒置显微镜下观察细胞的生长状态,用细胞水通道蛋白5(AQP5)、肺表面活性物质相关蛋白(SPC)、植物凝集素(BSI)、波形蛋白(Vimentin)4种肺细胞的特异性表达产物鉴定细胞。结果 接种2 d时,细胞开始贴壁生长。4~6 d时细胞开始增殖,呈现梭形、立方形、多角形等多种形态。8 d时细胞增殖能力达到最大,细胞活力为(87±8)%,细胞纯度为(87±5)%。结论 对组织取材、分离和培养进行改进可获得产量高、生长状态好、纯度高的ATⅠ。
英文摘要:
      【Abstract】 Objective To improve the method of culturing and obtain purified alveolar epithelial typeⅠ (ATⅠ) cells of SD rats. Methods 1 d newborn SD rats were applied for cell culture and brains were decapitated for lung tissues obtaining after respiratory cessation. CollagenaseⅠand DNaseⅠ were used to digest and isolate cells. Then, cells were put into the plate coated with typeⅠ rat tail collagen and different adherence was used to purify cells. Meanwhile, culture medium replacement was conducted after 48 h. The growth status was observed under an inverted microscope. Immunofluorescence including specific marker AQP5, SPC, BSI, Vimentin were used to identify cells. Results 2 d after incubation, the cells began to adhere to plate. At day 4 and 6, cells began to proliferate and exhibited a shape of spindle, cube and polygon. 8 d after incubation, the character of proliferation reached the highest and the cell viability was (87±8)% and purification was (87±5)%. Conclusion By modifying the methods of tissues harvest, isolation and culture, we can obtain AT Ⅰ cells with high yield, good growth state and super purity.
查看全文  查看/发表评论  下载PDF阅读器
关闭