z****u 发帖数: 1007 | 1 我觉得在identify Pax7 + satellite cell 之后,之前的很多myoblast cell culture
protocol可能需要re-consider了。比如上面那个protocol,neonatal mice limb
muscle.分离出来的很有可能是混杂了satellite cell以及committed myoblast cell的
mixture,而到底是哪种细胞长起来就很难说了。
adult的myoblast cell culture好像没有吧。 我没有仔细查过文献,只是大概的尝试
过几次,觉得从adult muscle fiber分离出来的细胞体外很难长。 |
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f*****f 发帖数: 195 | 2 就实验研究手段而言,两者有啥区别?
satellite cell可以认为是muscle stem cell,我看到paper上大部分都是小鼠in vivo
的实验。primary myoblast实际上也是将satellite cell传代分离得到的。那么,假如
体外培养muscle stem cell,不就变成了myoblast?
谢谢! |
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m******5 发帖数: 1383 | 4 Sorry my major is intracellur trafficking, not familiar with myoblast before.
I actually read a lot of papers related to this period(later mitosis,G0--G1)
. I knew before many important events happened here.
But I am not sure if it is because a 'context' easy to manipulate and handle
or because it is 'really important' |
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z****u 发帖数: 1007 | 5 不是很明白你这个问题。
传统说法是Pax7+ satellite cell变成myogenic cells, proliferate.. primary
myogenesis
then secondary myogenesis, myoblast cluster and form the myotubes.
体外养satellite cells的方式好像很局限。我还没看到好的protocl能体外扩增
satellite cell without losing the undifferentiate status.
vivo |
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发帖数: 1 | 6
Download pdf published by SfN (Chapter from 'The History of Neuroscience in
Autobiography, Volume 8' edited by Larry R. Squire)
Yuh-Nung Jan's CV
Lily Jan's CV
Yuh-Nung Jan and Lily Jan
Birth
Family History and Growing Up
National Taiwan University
The Hiking Trip to Shitou in the Spring of 1967
Graduate School Application
Graduate Study at Caltech (1968锟974)
Seymour Benzer Lab (1974锟977)
Steve Kuffler锟絪 Lab at H... 阅读全帖 |
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w******y 发帖数: 4871 | 8 NOVEL THERAPIES
Gene therapy — The identification of the gene responsible for the
dystrophinopathies led to initial excitement that transfer of a functioning
dystrophin protein (whether by transplanted myoblast or direct genetic
manipulation) may provide substantial benefit to or even "cure" affected
patients. Although myoblast transfer has been attempted in humans, the
results have thus far not been encouraging [47]. However, in one patient who
received a bone marrow transplant for severe combi... 阅读全帖 |
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m******5 发帖数: 1383 | 9 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********k 发帖数: 2818 | 11 It is way too weak, u can simply read Thomas Rando's review or many other
reviews, then u would have an
idea about it.... |
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m******5 发帖数: 1383 | 12 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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w********r 发帖数: 1431 | 13 你的primary myocyte是说satellite cell么?
如果是的话,我用lipoRNAiMax往primary myoblast里转过siRNA,效率还可以,我做
western看过。
转plasmid确实是进不去,我都是用Adenovirus。
不过看业内大牛的paper也有往里面裸转质粒的,具体用的啥试剂记不清楚了,你可以
去查查Michael Rudnicki的paper. |
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s******s 发帖数: 13035 | 14 应该不是MSC吧?fibro上头是啥啊?
另外,fibroblast和myoblast有啥关系?
俺是老鼠盲,查了半天觉得两个ms没啥关系,前面一个问题怎么也查不到 |
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b****r 发帖数: 17995 | 15
myoblast也是胚内中胚层来的,更多的不记得了,google吧 |
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w********r 发帖数: 1431 | 16 in the case of beta cell, I think it is more than "literally" argument ...
Existing beta cell dublication and progenitor cell differentiation are
totally two concepts. For the name of muscle satellite cell/primary myoblast
/muscle stem cell, we are also confused. So we call them "muscle progenitor
cell" in our recent paper... lol
exist
work
correct one here though), on muscle satellite/stem cells, about 6 ys ago,
one prominent stem cell biologist(I am sure that many on this board knows
him) we... 阅读全帖 |
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n********k 发帖数: 2818 | 17 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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y****i 发帖数: 2194 | 18 primary myoblast is not that difficult to make and way better than C2C12. |
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f*****f 发帖数: 195 | 19 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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l*******i 发帖数: 153 | 20 iPS本身最大的创新可能是1)技术的简易化,使得其迅速普及;2)其本身机制的研究
促进了epigenetics等领域的发展。
reprogramming这个概念早就存在。就体细胞向ESC逆编程而言,SCNT应该比iPS更好,
可惜的是,到目前为止还无法在人的细胞上完全实现。
iPS细胞刚出来时,几乎所有人都对其临床应用充满了期待,随着研究的深入,该领域
内怀疑其clinical therapy价值的人渐渐增多,其主要弱点有:
1)iPS与ESC的identity:Yamanaka现在似乎仍然坚持iPS与ESC是非常相似的,但是他
本人也承认现在iPS还根本无法取代ESC;相反的一派认为iPS与ESC根本就是两种不同的
细胞,无论是在transcriptome水平,还是epigenetic水平——蛋白组水平的研究,我
只看到两篇文章,不知为啥这方面的研究到目前还少之又少。当然,就目前已有的ESC
株,不同实验室分离的,不同人种来源的,相互之间也有比较大的差别,甚至这种差别
影响到了其pluripotency/分化能力。我个人的看法是,ESC是一个动态变化的cell
population(... 阅读全帖 |
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l**********1 发帖数: 5204 | 21 >ESC是一个动态变化的cell
population(或者说其heterogeneity非常大),即使subcloning以后,一个cell pool
里,各个细胞之间差别可能比我们想象的要大———无论是transcriptome水平,还是
epigenetic水平——但这一点,估计得靠singel cell analysis来验证的。
Sure
plus Monte Carlo Markov Chain Stochastic approch
pls refer
Hanna J et al. (2009)
Direct cell reprogramming is a stochastic process amenable to acceleration.
Nature. 462: 595-601.
link:
http://www.ncbi.nlm.nih.gov/pubmed/19898493
or
Max Flöttmann et al. (2012)
A Stochastic Model of Epigenetic Dynamics in Somatic Cell R... 阅读全帖 |
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w********h 发帖数: 12367 | 22 【 以下文字转载自 Biology 讨论区 】
发信人: lingzhuzi (), 信区: Biology
标 题: Re: 2012年诺贝尔生物医学奖
发信站: BBS 未名空间站 (Mon Oct 8 15:55:54 2012, 美东)
iPS本身最大的创新可能是1)技术的简易化,使得其迅速普及;2)其本身机制的研究
促进了epigenetics等领域的发展。
reprogramming这个概念早就存在。就体细胞向ESC逆编程而言,SCNT应该比iPS更好,
可惜的是,到目前为止还无法在人的细胞上完全实现。
iPS细胞刚出来时,几乎所有人都对其临床应用充满了期待,随着研究的深入,该领域
内怀疑其clinical therapy价值的人渐渐增多,其主要弱点有:
1)iPS与ESC的identity:Yamanaka现在似乎仍然坚持iPS与ESC是非常相似的,但是他
本人也承认现在iPS还根本无法取代ESC;相反的一派认为iPS与ESC根本就是两种不同的
细胞,无论是在transcriptome水平,还是epigenetic水平——蛋白组水平的研究,我
只看到两篇文章,不知为啥... 阅读全帖 |
|