w*******r
发帖数: 23 | 1 主要想研究stem cell和somatic cell之间的homeostasis。
一些文章报道说如果somatic cell受伤的话，就会发出信号，诱导stem cell增殖分化,
替代掉损伤的细胞。现在有没有建立起在体外模拟这一过程的系统？
如果要建立这一系统需要考虑哪些问题？
我觉得这一系统的好处就是方便在体外分离纯化相关的因子。 |
i*********0
发帖数: 915 | 2 会不会是stem cell自己受点小伤就开始增殖呢.
楼主说的,是不是就是验证in vivo的feedback的问题. |
c******r
发帖数: 3778 | 3 实验嘛，不行就自己发明啦。
比如可以先回答cell contact是否required for somatic cell induced stem cell
proliferation.
可以把somatic cell先做你想做的处理，然后放在culture里，跟stem cell用0.3um的
膜分开。这样如果stem cell proliferation增加了，那说明是soluble factor，在
culture medium里面。可以尝试用不同的antibody block。这样可以找到是什么factor。
如果stem cell不增加，那么可以把stem cell用CFSE之类的marker标记了，和处理过的
somatic cell混养，这样如果刺激stem cell proliferation，那么说明cell contact
很重要。那么可以focus on cell surface molecules，试试不同的knock down，看看
有没有哪个能block这个过程。
当然，注意做好controls。
具体实验设计要看情况，我的意思是你这个问题很有意思，不能因为没有现成的
protocol而妨碍你回答这个问题。要自己想办法，要自己设计实验来回答问题。
化,
【在 w*******r 的大作中提到】
: 主要想研究stem cell和somatic cell之间的homeostasis。
: 一些文章报道说如果somatic cell受伤的话，就会发出信号，诱导stem cell增殖分化,
: 替代掉损伤的细胞。现在有没有建立起在体外模拟这一过程的系统？
: 如果要建立这一系统需要考虑哪些问题？
: 我觉得这一系统的好处就是方便在体外分离纯化相关的因子。
|
n********k
发帖数: 2818 | 4 I think the harder part is not to establish a system but its relevance to in
vivo...in the related research areas but haven't followed this line much...
and for most of part, it is kind of a 20 century approach...that said,
people/I would love to see a robust in vitro system which could be used to
model in vivo in a meaningful way...BTW, there are tons of advantages coming
with such a system if a relevance could be established/validated...
化,
【在 w*******r 的大作中提到】
: 主要想研究stem cell和somatic cell之间的homeostasis。
: 一些文章报道说如果somatic cell受伤的话，就会发出信号，诱导stem cell增殖分化,
: 替代掉损伤的细胞。现在有没有建立起在体外模拟这一过程的系统？
: 如果要建立这一系统需要考虑哪些问题？
: 我觉得这一系统的好处就是方便在体外分离纯化相关的因子。
|
m*p
发帖数: 226 | 5 It seems that this hypothesis was tested in hemotopeitic system in vivo, eg.
exposure to radiation, or transplantation. Some of the factors and
cytokines were up-regulated. |
w*******r
发帖数: 23 | 6 Thanks. Your reply is very inspiring!
factor。
contact
【在 c******r 的大作中提到】
: 实验嘛，不行就自己发明啦。
: 比如可以先回答cell contact是否required for somatic cell induced stem cell
: proliferation.
: 可以把somatic cell先做你想做的处理，然后放在culture里，跟stem cell用0.3um的
: 膜分开。这样如果stem cell proliferation增加了，那说明是soluble factor，在
: culture medium里面。可以尝试用不同的antibody block。这样可以找到是什么factor。
: 如果stem cell不增加，那么可以把stem cell用CFSE之类的marker标记了，和处理过的
: somatic cell混养，这样如果刺激stem cell proliferation，那么说明cell contact
: 很重要。那么可以focus on cell surface molecules，试试不同的knock down，看看
: 有没有哪个能block这个过程。
|
w*******r
发帖数: 23 | 7 Sorry, I don't understand what you mean by *relavance*?
in
..
coming
【在 n********k 的大作中提到】
: I think the harder part is not to establish a system but its relevance to in
: vivo...in the related research areas but haven't followed this line much...
: and for most of part, it is kind of a 20 century approach...that said,
: people/I would love to see a robust in vitro system which could be used to
: model in vivo in a meaningful way...BTW, there are tons of advantages coming
: with such a system if a relevance could be established/validated...
:
: 化,
|
n********k
发帖数: 2818 | 8 meaning whether the in vitro insight you gather will hold true in vivo...and
how robust the in vitro system could be...if it is very robust and highly
representative, and have the potentials to offer novel insights, then it is
very much valuable, otherwise it is hard to say...Taking cancer field as an
example, tons of the in vitro insights from earlier days turn out to be
misleading or in vitro artifacts...
【在 w*******r 的大作中提到】
: Sorry, I don't understand what you mean by *relavance*?
:
: in
: ..
: coming
|
n********k
发帖数: 2818 | 9 meaning whether the in vitro insight you gather will hold true in vivo...and
how robust the in vitro system could be...if it is very robust and highly
representative, and have the potentials to offer novel insights, then it is
very much valuable, otherwise it is hard to say...Taking cancer field as an
example, tons of the in vitro insights from earlier days turn out to be
misleading or in vitro artifacts...
【在 w*******r 的大作中提到】
: Sorry, I don't understand what you mean by *relavance*?
:
: in
: ..
: coming
|
c******r
发帖数: 3778 | 10 选择in vivo还是in vitro system除了大家都知道的优缺点之外要考虑的还有一个问题
，就是目的是什么。
如果是一个成熟的project，已经有很多positive的pilot experiments，有很多in
vitro的data，需要证明in vivo的relevance，那么确实选用动物实验会更好，更有意
义。
但是如果是一个摸索性的前期实验，根据成败来决定project的概念是否成立。那么可
能选用一个更简单，快速的办法更有效。这时候如果直接上老鼠，不但价格贵，程序复
杂，手续繁琐，时间太长。如果失败risk就比较大。而即使成功，由于没有很多in
vitro的test没法同时进行，浪费很多时间。
lz这个，我看还处于project的早期阶段，还不知道是否能够做下去。用个简单的in
vitro实验摸索一下，可能比较合适。等比如说系统建立了，证明某个因子是介导这一
过程的，然后去in vivo系统里面confirm，可能更好一些。
and
is
an
【在 n********k 的大作中提到】
: meaning whether the in vitro insight you gather will hold true in vivo...and
: how robust the in vitro system could be...if it is very robust and highly
: representative, and have the potentials to offer novel insights, then it is
: very much valuable, otherwise it is hard to say...Taking cancer field as an
: example, tons of the in vitro insights from earlier days turn out to be
: misleading or in vitro artifacts...
|
w*******r
发帖数: 23 | 11 Could you please show me some papers about what you said? Thanks.I didn't
find any.
eg.
【在 m*p 的大作中提到】
: It seems that this hypothesis was tested in hemotopeitic system in vivo, eg.
: exposure to radiation, or transplantation. Some of the factors and
: cytokines were up-regulated.
|
w*******r
发帖数: 23 | 12 Exactly, that is what I am thinking about.
Besides, I think another advantage of this in vitro system is that you can
isolate and purify related factors more easily by biochemical methods than
in vivo.
【在 c******r 的大作中提到】
: 选择in vivo还是in vitro system除了大家都知道的优缺点之外要考虑的还有一个问题
: ，就是目的是什么。
: 如果是一个成熟的project，已经有很多positive的pilot experiments，有很多in
: vitro的data，需要证明in vivo的relevance，那么确实选用动物实验会更好，更有意
: 义。
: 但是如果是一个摸索性的前期实验，根据成败来决定project的概念是否成立。那么可
: 能选用一个更简单，快速的办法更有效。这时候如果直接上老鼠，不但价格贵，程序复
: 杂，手续繁琐，时间太长。如果失败risk就比较大。而即使成功，由于没有很多in
: vitro的test没法同时进行，浪费很多时间。
: lz这个，我看还处于project的早期阶段，还不知道是否能够做下去。用个简单的in
|
