Rozita Nasiri added an answer:What is the right injection amount for SkBr3 cells in white ICR mice?
Is one time injection of cells enough to establish cancer in mice? is 1* 10^7 cells enough or not ? How detect cancer after injection and how determine that is our interested cancer which established in mice?
Thanks a lotFollowing
Prem Subramaniam added an answer:Do anti-cancer drug stimulate xenograft growth?
I used HT29 cells to generate xenograft, and treat the mice with tubulin inhibitor. Due to the solubility issue, I used 5% Tween 80 to formulate the compound. Very wired, the compound stimulated the tumor growth. When I don't use any formulation component, just PBS, of course the drug solubility is not good in this case, the drug actually inhibited tumor growth. Does anybody have a clue what is the reason for the opposite result?
It is attached:Following
Farzad Khanipour added an answer:Is there any Iranian fellow willing to collaborate with us on the animal studies of our nanodrug delivery system?
We have been working on an anticancer drug delivery system based on PLGA-b-PEG copolymer nanoparticles, and now are looking for colleagues who are expert in cancer animal models and related in vivo experiments. Our preferential tumor model is MCF7 breast cancer. Please contact me for further detailed information. Thank you.
I appreciate your interest on this project. Actually, at this stage our plan is to start the tumor efficacy studies of our delivery system on mouse breast cancer tumor models MCF7, MDA-MB-435 or MDA-MB-231. However, for Biodistribution and pharmacokinetic studies we would need to have breast cancer models of SD rats. I think we could build up a good collaboration at these stages of animal studies, if you are seriously interested and ready for a firm commitment.Following
Colt Egelston added an answer:Does anyone have a protocol for T cell isolation from a spontaneous mouse model of breast cancer, MMTV-PyMT ?
I have tried using Mitlenyi's Gentlemacs with type III collagenase at 37C for about 45 minutes-60 minutes and I am fairly sure I am over-digesting the T cells. After processing to single cell suspensions, cells have poor viability and Thy1.2 isolation does not really enrich the T cells. Additionally, surface markers like CD62L and CD3 are being shed it appears.
Thanks everyone. We currently think that Liberase treatment for 30 minutes is the gentlest and most effective option.Following
Matteo Bocci added an answer:Does anybody have an idea how to analyze tumors and metastasis in MMTV mouse models?
We are trying to see change in MMTV mouse with our gene knockout. Can anybody suggest how to proceed with it. I mean what would be the strategy to observe the changes in terms of number/size of tumor formation, metastasis.
thanks in advance
The best solution would be to take mice at different time point for the metastatic study, while it will be enough to anesthetize and measure with a caliper the dimension of the 10 mammary tumors. The MMTV-model should have palpable tumors by the 8th week of age. Form that time on it will take extra 7 weeks to reach a full blown adenocarcinoma. As for the metastases, you should be able to observe micrometastases already at 12 weeks, while macrometastases should be present by 16 weeks, or at least this is what we see in our lab.
For our KO studies we have tumor measurement once a week for at least 4 weeks and we either study the growth from an initial indolent stage to a carcinoma (8-12 weeks) or the progression to an adenocarcinoma with metastatic phenotype (11-15 weeks).
Given our ethical permit, we are not allowed to keep mice alive for more than 18 weeks, but already at 16 weeks the tumor burden is so big that sometimes mice cannot walk any longer as the tumors impede their movements.
Pedro Veliça added an answer:How mutated are the spontaneous breast cancers that develop In the MMTV-PyMT mouse strain?
In this model, carcinogenesis is induced in breast tissue by inducing the Polyoma Middle T antigen with the MMTV promoter resulting in tumors that develop over the course of 15-20 weeks that mimic the different stages of human breast cancer.
Does anyone have any idea of how mutated these tumors become during their development and if so, how does that compare to human breast cancer?
I've found some data showing how CD4 T cells play a pro-tumor role (at least in the FVB background) but I was curious about CD8 T cells. Interesting about the ER, PR HER2 dynamics - wondering how much of that is due to deregulated gene expression/epigenetics or accumulation of mutations.
Greetings from the East Coast of Sweden.
Junhua Mai added an answer:How can I separate 4T1 cells from mouse host cells?
4T1/Balbc model is an typical model for breast cancer, but 4T1 cells lack of specific markers for flow sorting. Is there any way I can distinguish the 4T1 cells from the mouse host cells? Thank you very much.
Hi Rajkumar and Go, really awesome suggestions, thank you so much for the kind advice. I will try the ideas.Following
Shaima Jabbar added an answer:Does anyone have information about immunodeficient mice?
I have done xenograft tumor, I used NCRNU-Fsp/sp athythmic mice model. I did not find any tumor growth even after 4 months. Does anyone have idea about which models are the best for this type of experiment.
Thank you so much for your time
Thank you so much Anuj, your information really useful.Following
Peter Schoenfeld added an answer: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 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?
when there are sufficient cells, you can use an Oroboros oxygraph (Gnaiger) for measuring cell respiration. Cell respiration reflects mostly the activity of the respiratory chain.
When you have access to a Searhorse device, you can measure oxygen consumption and glycolytic activity.
Selvi Durmus added an answer:Is it possible to get tumors from the human SUM52PE cells in mice?
If yes, how many cells? in SC or into mammary fat pads? w/wo matrigel? estrogen supplementation required?
Thanks a lot for your answer. Unfortunately, it is not possible for me to work with nude mice for the moment, but I will consider irradiation as an option.
Ridwan Ibrahim added an answer:Can anyone share with me how to identify brain/cerebral tumorigenesis in mice?
I want to know if there is a symptom that can suggest cerebral/brain tumorigenesis has occurred in mice.
Maybe behavioral changes or an article that says when cerebral tumorigenesis occur in mice
Thank you very much Anuj SinghFollowing
Xiaogang Feng added an answer:What is the total cell number of a C57 mice Popliteal lymph node ?
Hi , I need to know the total cell number of a C57 mice popliteal lymph node.
depends on my eperience, it could be around 350000, which is counted from alive cell from 4 weeks old mice without any infection! Good luck!Following
Joe Graymer added an answer:Who is carrying out preclinical research with Doxorubicin derivatives?
In our research, we synthesized in a straightforward way a very potent fluorine derivative of Doxorubicin exhibiting nM IC50s in the two cell lines that we preliminarily tested. It could be especially interesting as a second-line treatment option (active in Dox-resistant strains) with possibly reduced cardiotoxicity.
Unfortunately we lack the knowledge and/or research focus to carry out further studies.
Does anybody know of a research group which is carrying out preclinical research on anthracycline derivatives?
Answers are greatly appreciated
I just became aware of an article about a new Anthracycline anticancer antibiotic from an Streptomyces, by Wei Li et al, in The Journal of Antibiotics, Mar 2015, and also about PMID 25319239 by Braña et al, 2015, perhaps by looking at these papers you find names of those working in the fieldFollowing
Enzo Fabrício Ribeiro Nascimento added an answer:Can anyone provide me with a reference/link that fully describes with figures the development of ACF and their appearance/features by methylene blue?
ACF in Rat Colon Cancer Model with AOM?
Can any one provide me with a reference/link that fully describes with figures the development of ACF and their appearance/features by methylene blue? Also How long is the fixation time prior to staining with MB? In the literature the time ranges between immediate to 48 hours following fixation in formalin.
hi please send the question for another author, mr martinez in the same paper
Vladimir A. Kulchitsky added an answer:Is mammary gland tumor induction in Sprague-dawley rats with rat mammary tumor cell line LA7 reliable for antitumor activity assessment?
Induction of a mammary gland tumor in female Sprague-dawley rats by inoculation of rat mammary gland tumor cell line, LA7. I would like to get some advice : Is this model reliable for assessment of anti-tumor action? I got some references for this model.
1. Evaluation of cytotoxic and chemotherapeutic properties of boldine in breast cancer using in vitro and in vivo models (Drug Design, Development and Therapy 2014:8 719–733)
2. Induction of mammary gland tumor in female Sprague-Dawley rats with LA7 cells (African Journal of Biotechnology Vol. 9(28), pp. 4491-4498, 12 July, 2010)
I'm looking to find a reliable tumor model with cell-line as cost and time are limiting factors. Thank you for your attention.
Currently, during preclinical testing in vitro are two critical positions. Firstly, it is desirable to use human cell line culture. And secondly, it is necessary to apply the model of 3D cell culture. It may be linear or culture cells of the primary cell culture. Both of these requests are directed to the maximum approximation test conditions to natural conditions characteristic of a person (including a three-dimensional principle of cell populations at the level of the whole organism).
Please, try to open the link below.
Sushil Kumar Middha added an answer:For in vivo studies is it necessary that we should make plant extracts of pure solvents (ethanolic, aqueous etc)?
Anti cancer study in wistar rats after chemically inducing tumour with DMBA.
I was planning for an anti cancer study of my plant extracts with hydro ethanolic solvent(1:1), but i couldnt find any reference with mixed solvents. All the studies were those of pure solvents. Is there any technical error or something with this type of study?
I am not sure why are you not getting any reference on Hydro Alcoholic solutions. 70% alcohal itself mean Hydro Alcoholic solution.Following
Vasanth Siruvallur Murali added an answer:Has anyone tried doing a Hela cell xenograft in NOD/SCID mice?
I am having trouble growing tumors on NOD/SCID mice with HeLa Cells. Has anyone tried growing tumors with this cell line? There is considerable inconsistency I am observing with the tumor growth and would be glad if I could get any suggestions or help.
Thank you again Dr. Uzzaman.Following
Sean M Geary added an answer:What's the proper cell number of the EL-4 cells I should inoculate?
I have tried 2x106 and 5x106 cells of EL-4 to inoculate in C57 mice，but when I use 2x106 the tumor could not form and when I use 5x106 the tumor will disappear at day 7 or 8.what's wrong with the cells?
I have experience with EL4 and their more immunogenic version E.G7 (transfected with OVA) and I have found that these cells readily form tumors in C57 mice (8 - 10 wks of age) at challenges of 1x106 - 1x107 subQ. I think you have mycoplasma contaminating your cell line - have you checked for mycoplasma? There are kits available for this. If you find your cells are contaminated I would get a new batch of EL4 cells (ideally from ATCC).
If your cells are actually mycoplasma negative then, barring some other form of contamination or cross contamination with another cell line, I would think that some technical aspect of the tumor challenge is to blame. Make sure you challenge with cells that are in log-phase of growth ( not overgrown) and that they are >90% viable. Also challenge with cells that are suspended in PBS or media that contains no foetal calf serum since that could act as an immunogen.
Mark J Mondrinos added an answer:Which is the best way to put tumoral cells into brain slices?
We are establishing mouse brain organotypic co-cultures with tumoral cells.
So far I've tested matrigel to localiced the cells in the slice, but when it gets in contact with the humidity of the slice, the mixture of matrigel and cells starts to expand and the cells ended everywhere.
Does anyone know any technique or procedure to localize these cells and avoid the difussion of the cells?
It is difficult to answer your question without more details of how you set up the experiment, but if the section is sufficiently thick (I assume it must be 300 micron or thicker) then perhaps you can inject a tiny volume (say 1 or 2 microliters) directly into the slice. If you take a small needle (25 gauge) and shove it onto a 10 microliter pipette (fits well on eppendorf pipettes) then you can pipette as low as 1 microliter of cell suspension (very high density) accurately. If it works you should get localized pocket of cells where you inject. In that case they would interact with and perhaps invade the surrounding tissue more rapidly.
If you stick with the gel matrix delivery, find a way to position it on top of or adjacent to the slice using some kind of improvised well/spacer as described here: http://www.jove.com/video/50881/coculture-system-with-an-organotypic-brain-slice-3d-spheroid
Finn von Eyben added an answer:How can I evaluate mouse tumor volume equivalents in human?
Can someone help me about mice to human conversions in the field of chemotherapy? I observed approximately 14 cm3 prostate cancer tumors in my control group animals (mice). However I do not know how to compare this volumes with human tumors. Is there a calculation method for this purpose
I do research on tumor volume for DIL. Localized prostate cancer is most often multifocal, so tumor volume may be the volume of the dominant lesion or index lesion, or the total sum of cance. Most stuies refected on the tumor volume for the index lesion. In everal recent studies the tumor volume of the index lesionwas in median around 2 cm3. These results were bsed on histopathology afer radical prostatectomy. I dont think we have siilar series oof studis whethe tumor volume is te sum of all tumor lesions in patients with metastatic prostate cancer. I wrote a paper on testicuar germ cell tumors observed in a patients and transplanted to nude mice.We described Growth curves for the TGCT in the two environments. I think you need to give more information before I can give a better answer.Following
Michelle Klein Sercundes added an answer:Regarding p53 knockout mice, which cell population in bone marrow and spleen is Gr-1+ CD11B- and which is lin-ckit+sca1-CD34-CD16/32hi ?
the above two questions are found in p53-/- mice
I do not know for sure what you are looking for in the bone marrow and spleen of these animals their knockouts, but if you are doing only one screening of the population I advise not using the GR1 antibody, use the Ly6C and Ly6G antibodies that can give you more safety of displaying populations. And as their cells are GR1 + CD11b-, there are reports that there dendritic cells expressing GR1.Following
Hozeifa Mohamed Hassan added an answer:Is it possible to store mouse organs in a -70C° deep freezer for one month?
I want to preserve major mouse organ, like brain, liver, lung, heart, kindney, and spleen. It will contain certain organic dye. Is it possible to store mouse organ in deep freezer (-70C) for a month? What would be the best method?
I agree with both Marthe-Susanna and Richard, first you should snap freeze the tissues in liquid nitrogen (for about 2-3 days), then transfer them immediately into -70 or -80 freezer, it will be preserved for yearsFollowing
David Jamieson added an answer:Can anyone help me understand how doxycycline and tetracycline reaches xenografts in mouse in-vivo studies?
Doxycycline, a modified form of tetracycline is regularly used to induce gene expression or suppress gene expression in xenografted cells transfected with TET-ON or TET-OFF systems. It is usually provided by drinking water. Can anyone help me understand how will it reach the xenografted cells or how long will it take?
Actually curious to know how it's efficiency might be affected if the cells are injected directly into the mammary ducts. (mammary intra-ductal xenograft model)
Just want to add to the above, it might be worth checking the MDR1 status of your zenograft cells. I think that doxycycline might be a substrate for P-gp and while enough might be getting into the mouse it could potentially not accumulate in your zenograft cells.Following
Ilya Tsyrlov added an answer:Is there a difference between HC Matrigel and GFR-HC Matrigel in tumor Formation?
I am planning to use Matrigel for tumor induction in mice. However on Corning I found an HC Matrigel and Growth Factor Reduced (GFR); HC Matrigel. Both are used for tumor induction in imunocompromised mice however does anyone have any experience in using either and if there are any pros and cons in either of them?
Murine NSCLC cell models could become a most valuable preclinical tool, and it will be interesting to see whether these NSCLC models are also refractory to chemotherapy and/or radiation therapy as so frequently observed for NSCLC. The murine models for SCLC and especially MSCLC already have many of the features of human SCLC, including metastatic spread. One of the most important translational aspects of MSCLC will be to determine its radio- and chemosensitivity. If similar sensitivities to classical drugs like cis-platin, carboplatin, etoposide, and irinotecan are found as seen in the treatment of human SCLCs, the model might help to unravel the molecular mechanisms underlying the development of chemoresistance. Finding MSCLC relapses after therapy would be extremely important since human SCLC does invariably relapse with current tumor intervention therapies. This would make the MSCLC model invaluable for testing both classic cytotoxic therapies and targeted therapies as well as combinations thereof.Following
Siddik Uzzaman added an answer:Which lung and liver cancer cell lines do you find grow best in nude mice?
We have synthesized liposomes for siRNA delivery and aims to examine in vitro and in vivo. Which lung and liver cancer cell lines do you find grow best in nude mice?
A549 cell line for lung cancer and HCC cell line for liver cancer. These cell lines will work I guess.Following
Hossein Rahmani added an answer:Has anyone maintained APCmin/+ mice on RM1 (Company: Special Diets Services) maintenance diet instead of RM3 breeding diet?
Recently, there was a change in diet of the APCmin/+ mouse colony on B6 background. Instead of being fed the RM3 (highly nutritious) breeding diet, the APCmin/+ breeders were fed RM1 (low in nutrients) maintenance diet. Any ideas if the parents not being on a breeding diet would affect the severity of phenotypes of the pups, such as polyp formation?Following
Hossein Rahmani added an answer:What is the range value for b2m in rats?
I want to know what point or what value of b2m in rat one can say it is indicative of tumour presence in rats exposed to chromium.Following
Jean Paul Bourgault added an answer:What dose (mg) of 17β-ESTRADIOL pellet (60 day release) should I use to grow MCF-7 induced tumors in SCID-SHO mice from charles river?
We are growing these tumors for a passive antibody transfer experiment to study effect on tumor growth
thanks Saku, will keep this in mind for next timeFollowing
Amirali Bukhari added an answer:Will mammary adenocarcinoma cell line from a Fischer 344 (F344) rat form a solid tumor in any F344 rat after subcutaneous administration?
Will 13762 MAT B III cells (ATCC CRL-1666; mammary gland adenocarcinoma cell line obtained from a Fischer 344 rat) grow efficaciously in any syngeneic (F344) rat and form a solid tumor after subcutaneous implantation and how long may it take to form a visible or palpable tumor? Will they do the same in other rat strains, e.g. in Wistar rats?
About 1.5 milion of the cancer cells are to be used for a single inoculation site.
Given the syngeneic nature of your experimentation, I don't see you facing any problem with them forming tumors. You should be able to see palpable tumors as early as 15-20 days.Following
About Cancer Animal Models
A forum to address questions regarding cancer animal models.