I'm so glad you are attending.
We have to make time for a drink.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 1/22/20, 1:01 PM, "Core Rigor and Reproducibility (CoRRe) on behalf of
Bover,Laura" <email obscured> on behalf of <email obscured>> wrote:
Thanks for the clarification! Obviously I was thinking in my type of
experiments, trying to identify new markers upon treatment with certain agents.
We used to have a Core here for identification of cell lines, but I am not
sure of it is still operating.
Thanks!
And see you soon at ABRF meeting!
Laura
Laura Bover, PhD
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of Biomedical
Sciences at Houston
Member of the Scientific Review Committee 2 (former Clinical Research
Committee 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St Room 1SCR4.2021
Houston- TX 77054
T 713-563-3301 <tel:713-563-3301>
F 713-563-3276 <tel:713-563-3276>
<email obscured>
On 1/21/20, 11:22 PM, "Core Rigor and Reproducibility (CoRRe) on behalf of
Frances Weis-Garcia" <email obscured> on behalf of
<email obscured>> wrote:
Thanks Laura and sorry for the confusion.
What I am looking for is a way to tell cell line apart if they come
from the same parent. For human lines that are from different people, STR
profiles are perfect ... but for lines derived from the same original (parent)
line ... there is currently no easy way to tell the difference between parent
line A and children lines A1, A2 much less grandchild A1.3. Make sense ... If
it was a hybridoma, I would just sequence the CDRs.
A colleague of mine and I were thinking of inserting a "DNA barcode" in
the lines, probably via CRISPR so we can put in as innocuous a site as
possible, but before developing a solving I was really hoping one already
existed ... so I thought I would poll this group ...
All about cell line authentication ... one of the many building blocks
needed for RnR.
Frances
ZRC 1563
Office 646-888-2354
Mobile 516-967-0656
On 1/21/20, 7:39 PM, "Core Rigor and Reproducibility (CoRRe) on behalf
of Bover,Laura" <email obscured> on behalf of <email obscured>> wrote:
So, do you need to differentiate the parent cells from the ones
resulted from the division or proliferating?
May be is not your question.
What about staining with CFSE a bunch of cells and let them to
divide and sort the different peaks (usually 7, at least in normal cells such
as T cells proliferating) by FACS. The mean fluorescence as you know is going
lower with each division. Is that what you need?
Depending on the proliferation time you will be able to separate
the last progeny, as well as the intermediate.
In normal T cells after 5 days you can recover cells from each
division. Not sure if with a cell line you could do the same.
Laura Bover, PhD
Professor
Director Monoclonal Antibody Core Facility
University of Texas M.D.Anderson Cancer Center
Associate member of The University of Texas Graduate School of
Biomedical Sciences at Houston
Member of the Scientific Review Committee 2 (former CRC 2)
Member of the Steering Committee of the Immunology Program GSBS
Immunology Department/ Genomics Medicine Department
7455 Fannin St Room 1SCR4.2021
Houston- TX 77054
T 713-563-3301 <tel:713-563-3301>
F 713-563-3276 <tel:713-563-3276>
<email obscured>
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On 1/21/20, 8:15 AM, "Core Rigor and Reproducibility (CoRRe) on
behalf of Frances Weis-Garcia" <email obscured> on behalf of
<email obscured>> wrote:
WARNING: This email originated from outside of MD Anderson.
Please validate the sender's email address before clicking on links or
attachments as they may not be safe.
Question ... Does anyone know of a way to reliably
differentiate between decedents / children of a cell line as well as the
parent?
One solution is to insert a plasmid to serve as an internal
barcode, but maybe there are other options ...
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