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楼主 |
发表于 2018-3-30 00:53:51
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这是不连续密度梯度离心分离TIL的原始文献: Suspensions of disaggre-gated tumors (generally in 5 ml of medium) were placed on top of the gradient formed by overlay- ing a cushion of 100% Ficoll-Hypaque (density 1.077, Pharmacia, Uppsala, Sweden) with an equal volume of 75% (v/v) Ficoll-Hypaque in RPMI 1640 (density, 1.055). Gradients (25 ml) were centrifuged at 800 × g for 20 min at room temperature, The MNC-enriched fractions which formed a distinct band at the interface between 75% and 100% Ficoll-Hypaque were collected, washed twice in fresh medium and counted in a hemacytometer. Tumor cells, which formed the upper band on top of 75% Ficoll-Hypaque were also collected.
我可以这么翻译吗:离心管中先加入10毫升100% Ficoll,然后再加入10毫升用1640或者生理盐水稀释的75%Ficoll,最后在上面加入5毫升的肿瘤细胞悬液,800 g离心20分钟。这样肿瘤细胞就在最上层了,是75%Ficoll的上表面,TIL就是在75%Ficoll和100%Ficoll之间了是吗?
和附件中的图片所示一致。
800g速度太高,我一般是400g离心30分钟。 |
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