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9.7K Views
•
07:01 min
March 1st, 2020
DOI :
10.3791/60022-v
Chapters
0:05
Introduction
0:35
Sphere Formation and Derived Adherent Population Generation
2:28
Sphere-forming Capacity Determination
3:02
Self-renewal Determination
3:59
Sphere Projection Area Determination
4:51
Results: Representative Endometrial and Breast Cancer Cell Sphere and Derived Adherent Population Analyses
6:29
Conclusion
Transcript
该协议允许使用功能测定和表型表征,以稳健的方式识别和分离癌症干细胞。从肿瘤样本中分离癌症干细胞可以作为一个平台,指导特定疗法在单个肿瘤的临床应用,预测耐药性和随后的复发。为了准备非粘附悬浮培养物,首先将细胞培养容器涂上50微升,每毫升聚HEMA为15毫克,然后将容器放入37摄氏度的干燥炉中至少两天。
当容器完全干燥时,用 PBS 清洗 80% 至 90% 的汇合癌细胞系培养,然后用 1 到 2 毫升的三辛 EDTA 分离细胞。在37摄氏度下5分钟后,用2到4毫升的新鲜细胞培养所
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Summary
该方法的目的是使用功能测定和表型表征特征,通过流式细胞学和西式细胞测定法,以强有力的方式,用球形形成方案识别癌细胞系和原发性人类肿瘤样本中的癌症干细胞(CSC)。杂交。
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