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Question:
4T1 metastatic breast cancer model which is a syngeneic xenograft model based on 4T1-12B - can anyone help?
Can the 4T1 metastatic breast cancer model - which is a syngeneic xenograft model based on 4T1-12B - be used as a model to study the anticancer effect...
[more]
Can the 4T1 metastatic breast cancer model - which is a syngeneic xenograft model based on 4T1-12B - be used as a model to study the anticancer effects of plant based compounds without injecting into a balb/c mice? Can one culture it directly in flasks, apply plant based extracts and study the effects of plants on the cells?
By Stephen Karori
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Egerton University
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Question:
Isolating CD3+ T cells from mouse tumor tissue (C57LB/6 and BALB/c) - MACS Beads
I am injecting B6 and BALB/c mice with B16F10 and 4T1 cells, respectively, to generate melanoma and breast cancer models. I then want to extract the t...
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I am injecting B6 and BALB/c mice with B16F10 and 4T1 cells, respectively, to generate melanoma and breast cancer models. I then want to extract the tumors, digest in Collagenase IV/DNase I and then centrifuge on a Ficoll column to isolate lymphocytes. Next I want to grab all the T cells, and since our lab has the MAC Beads system from MiltenyiBiotec I want to get some CD3 beads from there.
I am quite confused on what are the best beads to order. It seems human samples can use the CD3 MACS beads, but mice aren't as straightforward. I want to isolated, not deplete, CD3 cells, so it seems the CD3e beads on the website wont be applicable to my experiment.
In your guys opinions, what are the best beads to order?:
Pan T Cell Isolation Kit II
https://www.miltenyibiotec.com/Products-and-Services/MACS-Cell-Separation/Cell-separation-reagents/T-cells/Pan-T-Cell-Isolation-Kit-II-mouse.aspx
CD90.2 Microbeads
https://www.miltenyibiotec.com/Products-and-Services/MACS-Cell-Separation/Cell-separation-reagents/T-cells/CD90-2-MicroBeads-mouse.aspx
Those are the only 2 I found that can isolate CD3+ T cells, I don't want to separate CD4/CD8 from them, I just want all possible T cells to be used with one column. Separation into CD4/CD8 will later occur using cell sorting with the FACSAria III.
By Nathan Johnston
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The University of Western Ontario
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Question:
4T1 metastatic breast cancer model which is a syngeneic xenograft model based on 4T1-12B - can anyone help?
Can the 4T1 metastatic breast cancer model - which is a syngeneic xenograft model based on 4T1-12B - be used as a model to study the anticancer effect...
[more]
Can the 4T1 metastatic breast cancer model - which is a syngeneic xenograft model based on 4T1-12B - be used as a model to study the anticancer effects of plant based compounds without injecting into a balb/c mice?
Can one culture it directly in flasks, apply plant based extracts and study the effects of plants on the cells?
By Stephen Karori
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Egerton University
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Question:
What are the most suitable breast cancer cell lines and colorectal cancer cell lines for orthotopic NUDE mouse models?
I am looking for the preferred best suitable breast cancer cell lines & colorectal cancer cell lines for orthotopic NUDE mouse. Please share your exp...
[more]
I am looking for the preferred best suitable breast cancer cell lines & colorectal cancer cell lines for orthotopic NUDE mouse. Please share your experiences about the cell line number and other relevant details.
By Milind Sagar
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Sphaera Pharma
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Question:
Hind limb ischemia in SCID mice?
We are facing a problem in induction of hind limb ischemia in SCID mice. We tried common femoral artery ligation (only knot) and also cut, but it is n...
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We are facing a problem in induction of hind limb ischemia in SCID mice. We tried common femoral artery ligation (only knot) and also cut, but it is not inducing ischemia as expected. The limb is totally functional, no difference in the muscle mass and the histo-pathology also showed negative results.
Interesting thing here is, when we tried the same surgical methods in BALB C mice, which is the background strain for the SCID mice, the surgery worked perfectly well and the limb lost its function completely. Histo-pathology results were also positive in BALB mice.
So can anyone suggest the best surgical method for inducing hind limb ischemia in SCID mice? Please cite any references.
By Mallu Abhiram Charan Tej
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Indian Institute of Technology Madras
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Question:
Can someone answer some questions regarding rat / mouse serum / plasma measurements?
I am interested in measuring insulin and nor-epinephrine levels in rodents (rat / mouse).
What are the best methods or kits available, which use the l...
[more]
I am interested in measuring insulin and nor-epinephrine levels in rodents (rat / mouse).
What are the best methods or kits available, which use the least possible amounts of serum /plasma/blood to do the same?
Are there any arrays to quantify cytokines and hormones from various body fluids of rodents (blood / serum / plasma / urine)?
By Mallu Abhiram Charan Tej
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Indian Institute of Technology Madras
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Question:
What is the cell cycle profile in DT40 cells?
I was told normally the doubling time is 8 hours at 39C and that 80% of the population should be in S-phase. How long does the cell spend in each cell...
[more]
I was told normally the doubling time is 8 hours at 39C and that 80% of the population should be in S-phase. How long does the cell spend in each cell cycle phase? Thanks for reading.
By Shih-Chieh Chiang
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University of Sussex
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Question:
Control cell lines for drug synergy experiment?
I am developing combinations of targeted drugs for cancer therapy.
In order to make sure that the effective combinations of drugs I select in vitro w...
[more]
I am developing combinations of targeted drugs for cancer therapy.
In order to make sure that the effective combinations of drugs I select in vitro will not turn out to be too toxic in vivo, I want include a few 'normal' cell lines as control. I expect the drugs IC50s to be significantly higher on the normal lines than the cancer lines I use.
So, what would be the best 'normal' control to include in my experiments? Would the usual 3T3 be sufficient ? Or should I tailor the choice of control lines (e.g. of different tissue origin) based on the known in vivo toxicities of the drugs I choose?
By Lorenzo Bombardelli
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Netherlands Cancer Institute
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Question:
Are there any murine breast cancer cell lines derived from C57Bl6?
I've already used some human cell lines for in vitro assays, but I would like to inject cancer cells in C57Bl6 mice (not nude).
I've already used some human cell lines for in vitro assays, but I would like to inject cancer cells in C57Bl6 mice (not nude).
By Olivia Fromigué
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Institut national de la santé et de la recherche médicale
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Question:
What drives to predict the signaling cascade in response of a particular stimuli?
I just want to know how to predict the response of a cell that activated a signalling event. I wish to discover any fundamental or computational tool ...
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I just want to know how to predict the response of a cell that activated a signalling event. I wish to discover any fundamental or computational tool to predict the signalling events.
If any exist, by giving the search phrase to the web to predict the possible mechanisms at dependent experimental conditions?
By Srikanth Aluru
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Aix-Marseille Université
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Question:
Does anyone have experience determining human origin of cancer cells grafted to a mouse?
I'm looking for a reagent that will distinguish human from mouse cells by IHC-P
I'm looking for a reagent that will distinguish human from mouse cells by IHC-P
By Natasha Deane
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Vanderbilt University
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Question:
Paclitaxel dosage used in mice models.
Anyone knows the best/ mostcommonly used dosage of paclitaxel in mice to mimic cancer patients chemotherapy?
Anyone knows the best/ mostcommonly used dosage of paclitaxel in mice to mimic cancer patients chemotherapy?
By Geraldine Chiew
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Nanyang Technological University
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Question:
How can I develop a rat model for mammary tumours in normal rats without inducing the tumour chemically?
I followed a method for the development of mammary tumours in spargue dawley rats by using DMBA. According to the article, after 15-20 weeks for the t...
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I followed a method for the development of mammary tumours in spargue dawley rats by using DMBA. According to the article, after 15-20 weeks for the tumour growth could have shown. It is more than 30 weeks after the dose of DMBA and no tumour growth is observed. I could have the lesser average body weight of rats comparatively controls and vehicles after the treatment but no tumour induced till now. I want to induce mammary tumours in some of the rats transgenically or surgically or any other methods. Could anybody suggest something?
By Shankar Suman
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Indian Institute of Toxicology Research
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Question:
I am gonna inject MCF-7 cells to nude mices , do you have a suggestion for injection amount ? 1* 10^7 cells is enough or not ?
and should I add Extra Fetal bovine serum, antibiotics or fetal bovine insulin during injection ?
and should I add Extra Fetal bovine serum, antibiotics or fetal bovine insulin during injection ?
By Tuğba Kiriş
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Fatih University
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Question:
How can I turn the pLKO1 vector into a vector for protein over expression?
I would like to do gain-of-function and loss-of-function experiments for my gene of interest in a cell line (293T) first, and later xenograft these ce...
[more]
I would like to do gain-of-function and loss-of-function experiments for my gene of interest in a cell line (293T) first, and later xenograft these cells into mice and check for differences in tumour growth.
The knock down shRNA is in pLKO1 (which works fine) but the overexpression vector is pcDNA3.1.
Any possibility to overexpress the protein in the pLKO1 vector? That would save me one control group in the animal experiment.
Which modifications are necessary? Would the U6 promoter sufficient to drive also protein overexpression?
By Alexander Henke
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Universität Köln
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Question:
What is the best In Vitro assay? Alamar Blue, Presto Blue, XTT or MTT?
The Assay for in vitro activity..
The Assay for in vitro activity..
By Satishkumar Tala
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Academia Sinica
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Question:
Has anoyone injected human PBMCs in SCID mice?
How many cells do I inject and if in PBS or any thing else. which route of injection is preferred?
How many cells do I inject and if in PBS or any thing else. which route of injection is preferred?
By Mallu Abhiram Charan Tej
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Indian Institute of Technology Madras
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Question:
What could be the reason of disappearing tumors after growing up to 100 mm^3 of PC-3 cell line (Prostate cancer) inoculated in CD1 male nude mice?
Can we use female CD1 Nude mice also for tumor growth of PC-3 Cell line? Are CD1 nude mice species specific for certain cell lines only?
Can we use female CD1 Nude mice also for tumor growth of PC-3 Cell line? Are CD1 nude mice species specific for certain cell lines only?
By Milind Sagar
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Sphaera Pharma
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Question:
Open
Can we develop breast cancer cells (MCF-7) in swiss albino mice?
Due to lack of nude mice we are interested to develop breast cancer in normal mice. does any one have the idea?
Due to lack of nude mice we are interested to develop breast cancer in normal mice. does any one have the idea?
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Question:
Does anyone have experience in setting up primary cultures of epithelial cells from Sprague Dawley rat mammary tumors induced by DMBA?
I have used collagenase II and trypsin for tissue disruption and cultures in DMEM/F12 with 10% of fetal bovine serum but I get a lot of fibroblast con...
[more]
I have used collagenase II and trypsin for tissue disruption and cultures in DMEM/F12 with 10% of fetal bovine serum but I get a lot of fibroblast contaminating the culture.
By Corina Sasso
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National Council of Scientific and Technical Research
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Question:
How do I reduce the self-fluorescence of cells of Ehrlich ascites carcinoma while performing an assay of DCFH-DA, in flow cytometry?
The papers we assessed don´t explain about this parameter, and I´d like to know if only compensation is the solution for the problem.
The papers we assessed don´t explain about this parameter, and I´d like to know if only compensation is the solution for the problem.
By Monalisa Brito
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Universidade Federal da Paraíba
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Question:
Is it possible to measure the activity of the mitochondrial respiratory chain in vivo?
We have reasons to think due to metabonomic analysis that different tumors show different glycolytic / respiratory rates. There are a lot of technique...
[more]
We have reasons to think due to metabonomic analysis that different tumors show different glycolytic / respiratory rates. There are a lot of techniques for directly measuring media acidification (translating glycolysis) and oxygen consumption (translating mitochondrial activity) for cell culture, however I am not aware about how to perform such measurements in tissue in vivo - other than carbon tracking. Does anybody have any experience in this topic?
By Miguel Quintela-Fandino
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Centro Nacional de Investigaciones Oncológicas
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Question:
How to determine the therapeutic endpoint in an orthotopic pancreactic cancer model?
I want to measure the effect of a vaccine on the tumor burden in a syngeneic orthotopic pancreatic cancer model. We use Panc02 cells in C57Bl/6 mice f...
[more]
I want to measure the effect of a vaccine on the tumor burden in a syngeneic orthotopic pancreatic cancer model. We use Panc02 cells in C57Bl/6 mice for establishing the tumors. We tried using firefly luciferase expressing cells, but the signal we get from IVIS imaging is very inconsistent. My question is how do I determine when to sacrifice the mice and collect the tumors? Many papers do survival studies with orthotopic models and their endpoint is 200 mm2 tumor size, but they do not mention how they measure the tumor size. So, is it possible to use any other parameter for determining the endpoint? Like weight loss or appearance of distress?
By Shailendra Tallapaka
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University of Nebraska Medical Center
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Question:
What are the best practices for the taking and transport of tumors to be implanted as xenografts in mice?
We are thinking about the installation of a new research laboratory which would use xenografts for cancer therapy. The location of this laboratory is ...
[more]
We are thinking about the installation of a new research laboratory which would use xenografts for cancer therapy. The location of this laboratory is important for cost reasons. What are the best methods for taking and transporting tumors to be implanted in mice? Are there articles in the literature which would present best practices?
By André Martin
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École Polytechnique Fédérale de Lausanne
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Question:
Are any zebrafish researchers interested in collaboration on evaluating the function of Ikaros mutants?
We have data to suggest that mutant Ikaros cause lymphoid proliferation in Zebra fish. We would like to extend our studies to Ikaros mutants that are...
[more]
We have data to suggest that mutant Ikaros cause lymphoid proliferation in Zebra fish. We would like to extend our studies to Ikaros mutants that are resistant to SYK induced activation.
By Fatih Uckun
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University of Southern California
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Question:
A549 lung metastasis model
Has anybody done the tail vein injection of A549 for the lung metastasis? If so, what is the condition like and which animal do you use?
Has anybody done the tail vein injection of A549 for the lung metastasis? If so, what is the condition like and which animal do you use?
By Mayuko Omori
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Aragen Bioscience
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Question:
Is it better to analyse tumor infiltrating lymphocytes after isolation or to expand them with IL2 in vitro?
We have just started to work with tumor models, so maybe my question is basic. I want to isolate tumor infiltrating lymphocytes, but the thing I don't...
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We have just started to work with tumor models, so maybe my question is basic. I want to isolate tumor infiltrating lymphocytes, but the thing I don't understand is that in some papers some people use TIL right after isolation for FACS analysis and some expand TILs in vitro with IL2. What is the rational behind each approach? Isn't it better to analyse TILs right after isolation? In vitro expansion may influence results? Or am I wrong? Thank you for your help and for sharing your opinion!
By Adriana Tomic
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University of Rijeka
