B****w
发帖数: 48 | 1 不好意思只有一个包子的伪币了，请各位大侠原谅。
刚接触干细胞和免疫细胞，遇到一个问题。我把IPS细胞加药处理后，细胞好像分化为
了免疫细胞--看起来像巨噬细胞和T细胞。已经测了F4/80,CD11b，CD4,CD8，lysozyme
M.不知道进一步怎么证明他们就是巨噬细胞和T细胞。功能？还是有什么特异性的分子
标记？各位大侠可否给提供一两篇文献。十分谢谢！ | B****w
发帖数: 48 | | s****9
发帖数: 932 | 3 If they express T cell receptor (e.g. CD3 or TCRbeta), they are bona fide T
cells. You can stimulate them with plated bounded CD3/CD28 to see whether
they can proliferate, which is a function indicator for T cells.
Regarding macrophage, you could stain a few typical macrophage marker, such
as F4/80, CD11b, lysozyme M as you mentioned. The morphology of macrophage
is quite different from T cells as well.
You do not need a reference. Instead, you should add some splenic T and
macrophage as positive controls.
For beginner in the field, the positive and negative controls are the keys
for your experiment success.
lysozyme
【在 B****w 的大作中提到】
: 不好意思只有一个包子的伪币了，请各位大侠原谅。
: 刚接触干细胞和免疫细胞，遇到一个问题。我把IPS细胞加药处理后，细胞好像分化为
: 了免疫细胞--看起来像巨噬细胞和T细胞。已经测了F4/80,CD11b，CD4,CD8，lysozyme
: M.不知道进一步怎么证明他们就是巨噬细胞和T细胞。功能？还是有什么特异性的分子
: 标记？各位大侠可否给提供一两篇文献。十分谢谢！
| B****w
发帖数: 48 | 4 太谢谢了，包子奉上。我的问题主要是怕人家不相信我得到的细胞是免疫细胞。
T
such
macrophage
【在 s****9 的大作中提到】
: If they express T cell receptor (e.g. CD3 or TCRbeta), they are bona fide T
: cells. You can stimulate them with plated bounded CD3/CD28 to see whether
: they can proliferate, which is a function indicator for T cells.
: Regarding macrophage, you could stain a few typical macrophage marker, such
: as F4/80, CD11b, lysozyme M as you mentioned. The morphology of macrophage
: is quite different from T cells as well.
: You do not need a reference. Instead, you should add some splenic T and
: macrophage as positive controls.
: For beginner in the field, the positive and negative controls are the keys
: for your experiment success.
| s****9
发帖数: 932 | 5 你如果有具体的结果或者疑问，我们可以线下联系。
【在 B****w 的大作中提到】
: 太谢谢了，包子奉上。我的问题主要是怕人家不相信我得到的细胞是免疫细胞。
:
: T
: such
: macrophage
| n***3
发帖数: 663 | 6 For functionality determination of macrophages, you can stimulate the cells
with Toll-like receptor agonists (e.g. LPS). The stimulated macrophages will
release TNF and IL-6. Also, macrophages are known to engulf foreign
pathogens, you might want to also verify their engulfing ability by using
certain bacteria.
It really depends on how far you want to go and how satisfied the reviewers
would be.
Good luck!
T
such
macrophage
【在 s****9 的大作中提到】
: If they express T cell receptor (e.g. CD3 or TCRbeta), they are bona fide T
: cells. You can stimulate them with plated bounded CD3/CD28 to see whether
: they can proliferate, which is a function indicator for T cells.
: Regarding macrophage, you could stain a few typical macrophage marker, such
: as F4/80, CD11b, lysozyme M as you mentioned. The morphology of macrophage
: is quite different from T cells as well.
: You do not need a reference. Instead, you should add some splenic T and
: macrophage as positive controls.
: For beginner in the field, the positive and negative controls are the keys
: for your experiment success.
| B****w
发帖数: 48 | | w******y
发帖数: 4871 | 8 可不可以上flowcytometry啊，看看表达那些CDmarker或其它的marker,就应该知道是T,
B,NK,macrophage了吧 | B****w
发帖数: 48 | 9 上flowcytometry了，可是很多人说不是免疫细胞也可以表达他们的CD marker。人家不
相信我得到的细胞是免疫细胞。 | n***3
发帖数: 663 | 10 that's exactly why you need to do some functionality assay to further verify
these cells are indeed immune cells.
【在 B****w 的大作中提到】
: 上flowcytometry了，可是很多人说不是免疫细胞也可以表达他们的CD marker。人家不
: 相信我得到的细胞是免疫细胞。
| c******r
发帖数: 3778 | 11 还是做点immunohistochemistry吧，能看见的东西比较有说服力哈
当然功能性实验也很重要。
flow确实很好，很方便很快。但是不够直观。对很多pathologist来说还是看见才放心
啊。
【在 B****w 的大作中提到】
: 上flowcytometry了，可是很多人说不是免疫细胞也可以表达他们的CD marker。人家不
: 相信我得到的细胞是免疫细胞。
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