天津医药

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小细胞肺癌细胞株H446肿瘤细胞球培养方法的优化

王正岩1,2,李岩磊3,仇晓菲3   

  1. 1. 家庭住址
    2. 天津医科大学病理教研室
    3. 天津医科大学
  • 收稿日期:2010-06-29 修回日期:2010-09-13 出版日期:2011-01-15 发布日期:2011-01-15
  • 通讯作者: 王正岩

Optimization of the method to culture human small cell lung cancer cell line H446 tumor cell spheres

  • Received:2010-06-29 Revised:2010-09-13 Published:2011-01-15 Online:2011-01-15
  • Contact: WANG zheng yan

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摘要： 摘要 目的：优化体外培养小细胞肺癌细胞株H446肿瘤细胞球的方法。方法：利用干细胞无血清培养技术，观察H446细胞在下述不同培养条件下肿瘤细胞球的生长情况：接种于超低吸附及非超低吸附培养板，调整细胞密度至1×104/ mL、1×105/ mL、1×106/ mL 和1×107/ mL，采用针头吹打法和移液器吹打法将肿瘤细胞球制成单细胞悬液用以进一步传代。结果：（1）超低吸附培养板第二天形成明显肿瘤细胞球，非超低吸附培养板第十天出现肿瘤细胞球。（2）1×106/ mL组每个高倍视野下约有7个肿瘤细胞球，每个细胞球约由几十个甚至上百个细胞组成。（3）针头吹打法传代后肿瘤细胞球活性和增殖能力明显优于移液器吹打法。结论：选择超低吸附培养板、1×106/ mL密度接种和针头吹打法可有效分离肿瘤细胞球

关键词: 肺肿瘤, 癌, 小细胞, 细胞系, 肿瘤, 培养基, 无血清, 细胞培养技术

Abstract: Abstract Objective: To optimize the method to culture the tumor cell spheres in small cell lung cancer (SCLC) cell line H446 in vitro. Methods: Using the serum- free culture technology, we observed the growth situation of the H446 tumor cell spheres in the following culture conditions. The cells were inoculated to ultra low adherent plates and non-ultra low adherent plates. The cell density was adjusted to 1 ×104/ mL, 1×105/ mL, 1×106/ mL and 1×107/ mL. The cell spheres were blowed into single cells through micropipettor blow method and pinhead blow method in order to further passages. Results: （1）In ultra low adherent plates, the tumor spheres appeared clearly the next day, while in the tenth day in non-ultra low adherent plates. （2）At the concentration of 1×106/ mL, there were about seven spheres per high power field and dozens of cells even hundreds of cells for each sphere.（3）In the pinhead blow method, the tumor spheres in second generation showed better activity and proliferative ability. Conclusion: Chosing ultra low adherent plate, the inoculum density of 1×106/ mL and the pinhead blow method can separate tumor spheres effectively.

Key words: lungneoplasms, carcinoma small cell, cell line, tumor, culture media, serum-free, cell culture techniques, 细胞培养技术