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Open to questions.

Posted by mnscott11 on 26 Jul 2008 at 19:29 GMT

I would greatly appreciate any comments or questions regarding this manuscript. Please do not be shy.

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I have a general question about the paper

Alan1 replied to mnscott11 on 05 Nov 2008 at 17:04 GMT

I'm a graduate student, and I have problem to understand Fig 6A, (was this a parafilm embeded section, whole tumor ,or cell),and why Fig6A did not look like the classical IHC figure(ex"Fig 6F).

And ouestion is that is the virus insertion analysis the best way to study clonality ?(any alternative?)

Thanks many

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RE: I have a general question about the paper

mnscott11 replied to Alan1 on 22 Apr 2009 at 05:43 GMT

Hello Alan,

As stated in the figure legend, Figure 6A is an IHC staining of a single colony of cells while they're attached to a 96-well plate. The background is white because of the transmitted light, whereas the brown stain is ESA localization.

Figure 6F is a classical H&E stain of a paraffin embedded section.

Finally, by studying lentiviral insertions and inverse PCR of the insertion site is a classical method used to demonstrate clonality and has increasingly been used in hematopoietic stem cell biology. It is the only true way to demonstrate that progeny and a stem cell originated from the same cell, as visual inspection of "single cells" prior to injection is not definitive. There could always be another cell not in the field of vision, or without even inspecting cells visually, heterogeneity may be generated by 2 different cells stuck together. By demonstrating common lentiviral insertion sites, its pretty safe to say the 2 populations didn't arise from different cells.

I hope that helps.

Scott

No competing interests declared.
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RE: Open to questions.

ogires replied to mnscott11 on 07 Jan 2010 at 13:05 GMT

Dear Scott,

obviously, this is an excellent article. For the first time to my knowledge, you demonstrated the proportional increase of tumour initiating cells upon standard chemotherapy.
Hence, I am left with a very positive comment only.
Except maybe one little point: ESA, although being used in the past, is not the currently agreed name. EpCAM is rather used instead (See Baeuerle et al. Brit J Cancer, 2007).

Questions:
1. what is the current idea on the accumulation of CSCs upon chemotherapy? Are they omitted because they do only temporarily divide?
2. how would you, in principle, envisage to change standard therapy according to those findings?

Thanks for the contribution and congratulations.
Olivier Gires

No competing interests declared.
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