请问那位前辈用悬浮培养的方法富集过癌细胞系中的CSCs？

KINGxuridongshe

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1 a) M* r* d8 t6 n2000 是不是有点少，六孔板

kakapo

for non-adherent cell, you can directly culture it in suspending
5 N  [/ G; ]4 l& j7 Vfor adherent cell, you firstly make cells pellet. you do not disperse the cells pellet and directly culture the pellet. the cells will form cells clustered spheroid
, [' Z0 x% f  X) y1 S- nthe spheroid size depend on original cell number.  Your question is not that clear. please, post your question in detail.

KINGxuridongshe

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) p, F  m/ F' z1 T你好，能把你做的A549 tumor sphere图片给我看看吗？我最近在做这个但是不是很理想，有些问题如果你方便能请教一下吗？/ c, Z( G# a, U9 u2 o

kakapo

sorry, I do not have lung cancer cell line in hand. Well, the cell cycle length of A549 cell is approximately 36 hours, and is longer than other tumor cell line. time for forming sphenoid of A549 will be longer.
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to make cell sphere, if i did the work, I am going (1) to pellet 50000 cells in tip of eppendorf. Next, I will (2) discard PBS ( in last wash), (3) gently add 100 micro-liter FBS. (4) keep cells in eppendorf until pellet is floating. (5) transfer pellet to the 96 well plate.

zz091115

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做过

zz091115

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7 N. Q0 F2 L" B0 u0 Y4 J, v$ {不少 够了

zz091115

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没有 我主做原代 只是拿这个细胞养过看了一下

KINGxuridongshe

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5 z. x6 C- q8 v3 E你好，看你的帖子对于干细胞悬浮球培养有很深的了解，我现在的课题和肺癌干细胞相关，但是现在在实验过程中用A549成球一直不理想，和文章中悬浮球的样子和培养的时间都有出入，所以想请教一下，希望你能在百忙之中指点一二，

细胞海洋

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发一个新主题 把你的问题详细说明 我给你加高亮

zz091115

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! x, C6 w" f# p; u版主让你开个新帖子 然后你详细说明我再回答你吧