n********k
发帖数: 2818 | 1 C2C12 is both a very easy and very hard line to work with and I am not aware
any other better established line other than this one or one could go with
primary lines...
Anyhow, If one follows closely certain routines, this line is very easy--
-passage before 80-90% confluence with no local overgrowth...Overgrowth is a
huge problem with this line, after 3-5 times, the entire culture will be
changed...Also, the confluence etc matters a lot for differentiation...
In addition, there are at lea... 阅读全帖 |
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l**x
发帖数: 52 | 2 谢谢neverthink。你给的信息很全，很有用。你知道，可以从哪买到另一种C2C12吗？
另外，目前用这个细胞做siRNA实验也不是特别成功，siRNA从Dharmacon买的，应该质
量可靠，试了Qiagen hiperfect和Invitron lipofectamine RNAiMAX，效果都不怎么好
. 用过Qiagen hiperfect做过别的几株细胞，knockdown都很好。不知道是C2C12的原因
还是怎么的
aware
with |
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n********k
发帖数: 2818 | 3 不过，C2C12确实是一个heterogeneous population，即时low serum诱导分化，也总有
一小部分的细胞是不分化的。把这部分细胞拿出来，可以继续长。
one can repeat this several times, and then no cells can differentiate
anymore...another thing is even if one makes clones from the line, it would
behave the same...there are indeed something rather interesting here and
c2c12 has been so very useful in the field...if I have extra hands and funds
, I may do something with the lines besides my main direction in neural
stuff...have any interest to collaborate on... 阅读全帖 |
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m******5
发帖数: 1383 | 4 一个是技术上的，serum starving对 C2c12 differentiation有多大影响
另一个，刚开始做（从另一个方向涉及过来），发现很多人都很关心G0-G1这段时间发
生的事情，想问一下，为什么？
1 因为确实很重要，这段时间manipulation可以影响分化
2 因为已经有许多关于这时期分子机制成型的研究，容易编出story，循环 |
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m******5
发帖数: 1383 | 5 一个是技术上的，serum starving对 C2c12 differentiation有多大影响
另一个，刚开始做（从另一个方向涉及过来），发现很多人都很关心G0-G1这段时间发
生的事情，想问一下，为什么？
1 因为确实很重要，这段时间manipulation可以影响分化
2 因为已经有许多关于这时期分子机制成型的研究，容易编出story，循环 |
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n**8
发帖数: 221 | 6 请教有转染过 C2C12 和 10T 1/2 的大侠，一般分别什么转染试剂适用于这两种细胞？
不甚感激！ |
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n**8
发帖数: 221 | 7 谢谢回复！
oranger大侠用Mirus LT-1转过C2C12？效果如何？ |
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o*****r
发帖数: 156 | 8 I didn't work on C2C12 before, so I don't know about this particular cell
line.
My suggestion is to try both LT-1 and lipofectamine. |
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v*********d
发帖数: 382 | 9 invitrogen Neon Transfection Syste
C2C12 90%+ efficiency. Should also work for 10T1/2
Best part, you can get a 2week free demo |
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n*********n
发帖数: 95 | 10 我要分化C3H10T1/2 细胞到肌细胞，请问谁有好的Protocol。我们实验室原先只做
C2C12分化，用的是2%Horse Serum.不知道适不适用于C3H10T1/2细胞.
有文章用5-azacytidine诱导分化，不知哪个更好。谢谢！ |
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l**x
发帖数: 52 | 11 现在用的是C2C12细胞，经常一不小心就长满整个平板，ATCC说这个细胞株不能长满平
板，另外，这个细胞表达myosin regulatory light chain的磷酸化特别低，很难用
WESTERN测出来,用的是Cell Signaling的regulatory light chain S19的磷酸化抗体。
大家有什么好的细胞株推荐吗？特别是来自于skeletal muscle的。另外，对抗体有什
么推荐的最好了，谢谢 |
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w***e
发帖数: 269 | 12 I like L6 line better than C2C12. You can try that as well. |
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y****i
发帖数: 2194 | 13 primary myoblast is not that difficult to make and way better than C2C12. |
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n********k
发帖数: 2818 | 14 second this...that said, the LZ seems a newbie too and so established line
could have some advantage...
For C2c12 lines, one could just request from different labs around, I forget
whether the other line is from helen Blau's lab at stanford or not...they
have a c2 line which is even tricker...
For knock-down, I never tried in the line and I don't see the reason but I
guess u could try the same shRNAs in c2 together with some other line, see
what happens... |
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g*********5
发帖数: 2533 | 15 I passage c2c12 cells when confluence...
no confluence passage is it important for the further study?
thanks |
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f*****f
发帖数: 195 | 16 不是siRNA对老鼠细胞株效果不好，而是因为C2C12或primary myoblast转染效率偏低，
但也有人转染siRNA成功了的。
可以用病毒转染miRNA质粒，
或者脂质体转染后加质粒抗性标记筛选（如puromycin等）效果也很好。 |
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n********k
发帖数: 2818 | 17 I think C2c12 transfection efficiency is not that bad at all for functional
studies...one just has to be know the right assay/approach to determine the
efficiency of the knockdown...or figure out a protocol to do highly
efficiently siRNA transfection...in general, siRNA is a lot easier to be
transfected compare to plasmids... |
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b****s
发帖数: 148 | 18 nah..
once C2C12 differentiated, it will never re-enter cell cycle.
2% horse serum is good for inducing differentiation, but switching back to
higher % serum won't help growth anymore if myogenesis is in progress. |
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b****s
发帖数: 148 | 19 C2C12 needs to be passaged when under 40-50% conflu.
over 60% confluency will potentially lead to partial differentiation.. and
windup with less viable cells. |
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