Capillary cloning of primary human tumor cells: assay miniaturization for drug efficacy testing.
Capillary cloning of primary human tumor cells: assay miniaturization for drug efficacy testing.
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原发性人类肿瘤细胞的毛细管克隆：用于药效测试的测定小型化。
DOI：
10.1002/stem.5530070507
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
MurphyJr,MJ
中科院分区：
文献类型：
--
作者：
Peng,XE;Chen,GZ;Lu,YF;MurphyJr,MJ
传统的双层琼脂法克隆人类肿瘤细胞需要大量活的肿瘤细胞和14-21天的培养。这些先决条件经常限制其作为一种分析方法的效用。为了规避这些限制，减少实验所需的药物量，我们进一步发展和小型化了在玻璃微毛细管中克隆人类肿瘤细胞的实验。培养液为50 μl，含15000个有核细胞，置于975 mm毛细管中培养7天。50个连续肿瘤活检的结果表明，克隆效率在0.007%至1.0%之间，总体上成功克隆了88%的所有测试肿瘤，并且具有良好的线性生长关系和化疗敏感性。这种小型化的检测方法为药物功效测试提供了明显的优势，包括高克隆效率、小肿瘤样本和药物需求、更快的检测周转时间以及对试剂和培养箱空间的总体保护。
The conventional double‐layer agar method of cloning human tumor cells requires a substantial number of viable tumor cells and 14–21 days of culture. These prerequisites frequently limit its utility as an assay. In an attempt to circumvent these limitations and to reduce the amount of drug that is needed in the assay, we have further developed and miniaturized the assay in which human tumor cells are cloned in glass microcapillary tubes. Cultures consisted of 50 μl containing 15,000 nucleated cells in 975 mm capillary tubes which were incubated for seven days. The results from 50 consecutive tumor biopsies resulted in cloning efficiencies, ranging from 0.007% to 1.0% with an overall successful cloning of 88% of all tumors tested and a good linear growth relationship and chemotherapy sensitivity. This miniaturized assay offers distinct advantages for drug efficacy testing including high cloning efficiencies, small tumor sample and drug requirements, quicker assay turnaround time and a general conservancy of reagents and incubator space.