Questions related to Cancer Biology
This generalizable immunotherapy approach to cancer seems compelling:
Unlike many other immunotherapies, this one does not require pre-defining antigens.
Based on research from the Levy lab at Stanford, this method introduces a non-specific technique to attack cancers by injecting immunoenhancing agents (TLR9 and OX40) locally into the tumor site. In mouse models, these agents activated a robust, targeted anti-tumoral response.
Clinical trials were launched in 2019/2020.
What's the best way to track the clinical trials and see how this therapy work on humans?
I am starting working with E0771 cell line since I have to establish an orthotopic breast tumor model in c57 mice but I have no experience with this cell line so I would really appreciate any advice that you can give me.
In particular, I saw the ATCC website and they say that it is better to culture these cells in a t-75 corning flask, maintaining cultures at a cell concentration between 6 x 10^4 and 8 x 10^4 cells/cm2, is this true also for your experience? How many cells do you plate in a 75 cm2 flask?
Do they grow fast? How many times per week do you subculture them?
Sorry for all of these questions but I am new with these cells and so I would really be very grateful for all your advice,
Thanks a lot,
Cancer incidence is increasing globally. It is widely believed that increased life span is the main reason cancer risk overall is rising. A paper published in the Lancet reports that delays in screening, diagnosis, and treatment due to the COVID-19 pandemic could lead to excess cancer deaths, and slow or even reverse the declining trend in mortality projected for some cancers. https://www.thelancet.com/journals/lanpub/article/PIIS2468-2667(22)00111-6/fulltext
Furthermore, the report by Harvard Medical School researchers at Dana-Farber Cancer Institute and colleagues from other institutions, suggests that COVID-19 has complicated the treatment for patients with cancer. "In patients with cancer, COVID-19 can be especially harsh. This is likely because many of these patients have a weakened immune system—either as a result of the cancer itself or the therapies used to treat it—and are therefore less able to fight off infection by the new coronavirus". https://hms.harvard.edu/news/covid-19/cancer-interplay#:~:text=In%20patients%20with%20cancer%2C%20COVID,infection%20by%20the%20new%20coronavirus
In 2021, a research team led by Zhou highlighted the clinical and molecular similarities between cancer and COVID-19 and summarized the four major signaling pathways at the intersection of COVID-19 and cancer, namely, cytokine, type I interferon (IFN-I), androgen receptor (AR), and immune checkpoint signaling. They also discussed the advantages and disadvantages of repurposing anticancer treatment for the treatment of COVID-19. https://ijhoscr.tums.ac.ir/index.php/ijhoscr/article/view/1408
Jafarzadeh et al. reported that the patients with some types of cancers may be more vulnerable to SARS-CoV-2 infection compared with the non-cancerous individuals, due to their immunocompromised state resulted from malignancy, chemotherapy, and other concomitant abnormalities as well as perhaps greater expression of angiotensin-converting enzyme 2. Moreover, they reported that clinically recovered COVID-19 individuals display immune abnormalities that persist several months after discharge.
The lymphopenia-related immunosuppression, functional exhaustion of cytotoxic lymphocytes (such as CD8+ cytotoxic T-cells and natural killer cells), hyperinflammatory responses, oxidative stress, downregulation of interferon response, development of the myeloid-derived suppressor cells, downregulation of tumor suppressor proteins and perhaps reactivation of the latent oncogenic viruses may directly and/or indirectly play a role in the cancer development and recurrence in severe COVID-19 patients. https://ijhoscr.tums.ac.ir/index.php/ijhoscr/article/view/1408
I have a trouble need to solve for my experiment. I work in breast cancer cells. In my research, I want to check the adipocytic stimulation after treatment of Docetaxel and docxorubicin. As you know, cells will die after docetaxel and doxorubicin treatment. 72hours after anticancer drug treatment, I did Oil Red O staining for adipocytes stain. After that, i measured for OD at 500nm. But I want to normalize how many cells in each sample after staining. It means that I want to check how many cells were stained for Oil Red O stain. Does anyone have a suggestion?
Current search engines for MS/MS protein identifications such as: Mascot, MS Amanda, Sequest, etc., currently rely on the creation of a search library composed of computationally generated potential peptides through the cleavage by proteases (e.g., trypsin) of proteins from a given database. Different PTMs can be added to these computationally generated peptides, so that the search could be extended to address specific scientific questions, but this leads to significantly higher computational costs.
I have recently come across a case, where a highly enriched short protein could not be identified by a standard search, given that it was only generating a single peptide that had 2 fixed modifications. The modifications were not the most common there were and finding the right combination to use was time and computationally expensive.
I would like to open a discussion on the fact that pre-made peptidome libraries are a much better alternative to de-novo generated libraries of proteomes. Let’s get into the details!
As an example, I will use the ACE2 receptor, now infamously known to be the entry gate of Covid-19 into human cells.
The human ACE2 receptor undergoes a series of post translational event, such as: proteolytic cleavage by ADAM17 resulting in a soluble proteoform, glycosylation and phosphorylation of tyrosine-781 and serine 783.
In current search engines, the tryptic peptides generated would be generated from the first Methionine to the next positively charged residue and so on until the very last residue of the protein. If one would like to detect this protein in a sample and asses the presence of the mentioned PTMS, you would need to look for at least 2 phosphorylation sites per peptide and also check for S and Y phosphorylation. The search engine will then generate all possible combinations of SY single and double phosphorylate tryptic peptides to search for, which leads to exponentially increasing computational costs.
Since the protein is also cleaved by another protease in vivo, the 2 peptides before and after this site will not be accounted for as they do not end/begin after a positive residue. Since this is not a small protein, other peptides will probably still be detected, and the protein will eventually be identified.
I imagine a tool which would be used to generate the tryptic peptides as before, only accounting for the known PTM sites. In case of the ACE2 2 almost adjacent phosphorylation sites, this would lead to only 3 additional peptides (pY, pS, and pYpS). If the research question being asked is to identify novel phosphorylation sites, then only 1 phospho-site per peptide while looking for STY phosphorylation might already suffice, since the known ones will have already been accounted for. This can be applied to any combination of PTMs, massively reducing computational requirements. It is of course counterproductive to looking for PTMs in sterically inaccessible regions for example (e.g., hydrophobic core of the fold)
Databases of know annotated PTM sites of entire proteomes of many organisms are readily available. The tool could have a modular design in allowing the user to create a customized peptidome having any or all the following characteristics: trypsin/other enzyme used and/or accounting for known endogenous cleavage sites and/or accounting for known PTMs sites and/or accounting for natural variants.
I see a long list of advantages using this method and I would like to list the most important ones:
1. Identification of additional hits that could have been missed due to several reasons (e.g., tryptic peptides contain fixed modifications while not searching for these specific modifications due to computational resource limitation, or worse, small protein that would normally only yield in a single peptide that has 2 fixed modifications, one of which might be exotic)
2. Reduced computational time when trying to identify novel PTM sites
3. Lower false discovery rate since the peptidome used will be a much more closely related dataset to the actual sample composition than just a simple tryptic proteome and as a result newly identified spectra of interest can be more confidently assigned as the risk of artefacts is lower.
4. Single nucleotide polymorphisms can be analyzed analogously to PTM sites and would not result in exponentially larger search database.
5. More unique peptides could be assigned: If 2 proteins share a tryptic peptide, but one is known to be phosphorylated in this peptide but not the other, one could distinguish the phosphorylated peptide as having come only from one of the 2. In case of glycosylation this makes even more sense since some types of glycosylation only appear in a limited number of proteins, depending on their cellular localization
As the human proteoform project is taking on, maybe this would be the way of MS based proteomics to quickly catch up and help this project while advancing itself.
What are you thought on this? Are there any ongoing projects that would aim to do just that?
All tumors have DNA mutations, and a predictive understanding of those mutations could inform clinical treatments. However, 40% of the mutations are variants of unknown significance (VUS). So the challenge is to objectively predict whether a VUS is pathogenic and supports the tumor or whether it is benign. We are working on this problem and would welcome feedback on our efforts (see doi: 10.3389/fmolb.2021.791792 ) and also alternative ideas and insight.
I am doing my research on Cancer biology.
I need your suggestions in order to perform GWAS of autoghapy related gene in homo sapiens.
Kindly please suggest me some information about tools for the same.
Thank you all in advance.
I should use the BCH for inhibition experiment for LAT1. I saw that BCH is soluble in 1M NH4OH but I cannot use this solution with cells.
Could someone kindly clarify why aerobic glycolysis is required for cancer cells? That is, if tumor cells used OXPHOS, would they be unable to grow and metastasize as well?
The most commonly cited benefit of aerobic glycolysis seems to be accelerated glucose uptake, but why is this required for long-term growth as opposed to short-term spurts? The timeline for cancer progression is normally months or years, not hours or days.
Moreover, despite consuming glucose faster, total ATP production from aerobic glycolysis is lower. That is, during the time OXPHOS consumes 1 glucose molecule, aerobic glycolysis may consume 13 glucose molecules -- yet this yields fewer ATP molecules.
If correct, total energy production inadequately explains the relationship between cancer cells and aerobic glycolysis.
Is the timing of ATP essential? Perhaps cancer cells need fewer shots of ATP molecules but at a higher frequency rather than wait for one large batch from OXPHOS?
Or what are the other benefits of aerobic glycolysis over OXPHOS?
Ultimately, why would cancer cells be less successful with OXPHOS?
Thanks in advance for your help.
Metabolic rewiring and epigenetic remodeling, which are closely linked and reciprocally regulate each other, are among the well-known cancer hallmarks. Studies have reported use of Onco-metabolites to metabolically reprogram the epigenetic of cancer. I was wondering what might be major limitations of such techniques?
In the attached image, it shows the cleaved PARP densitometry analysis from western blot. Cleaved PARP is commonly used for apoptosis. Treatments (A and B) were tested along with control and the two time points were used 8h and 24h.
I'm trying to run qPCR experiment looking at mRNA levels of apoptosis markers such as BCL2 and PUMA. My question is do I choose 8h for qPCR assuming that mRNA comes before protein so looking at an earlier timepoint is better? Or do I choose 24h since I see the significance in cleaved PARP at 24h in western blot?
Iam interested in both Diversity and Cancer Genetics related Work... could u Please Suggest some research topic...it helps a lot...
I am trying to grow Kasumi-1 cell line. I use RPMI 1640 and 20 % FBS but cells never seem to be growing. They keep on shrinking in size and eventually die off.
This seminal paper by Paul Mischel rediscovered the relationship between extrachromosal DNA (ecDNA) and cancer:
1. How much would it cost to replicate the part of the experiment that generated subcutaneous tumors from seeds containing 200/2000/20000 cells of FACS-sorted EGFRvIII High/EGFRvIII Low subpopulations?
2. How much would it cost to sequence these tumor cells?
Thanks for your help!
The prevailing theory is that cancerous tumors reflect a form of cellular natural selection, where random mutations confer evolutionary advantages to tumor cells. These survival benefits accumulate over time, eventually empowering tumor cells to outcompete healthy cells and defeat the immune system.
Given this theory, one expects a correlation between cell division rate and cancer rate -- tissues with the highest number of cell divisions should show the highest incidence of cancer (environmental and hereditary factors notwithstanding).
However, few papers seem to verify this assumption, or even document cell division rates and lifetime cell divisions among different tissues.
These are the two best papers so far:
However, the same senior author leads both papers, and the papers seem to omit the most common cancer types, breast and prostrate. Furthermore, the methodology seems fragile as they conducted an analysis across different studies that may not have employed uniform methods.
Could anyone recommend more robust or authoritative papers on the subject of cell division rates?
Mitochondria can sense and control nutrient catabolism and anabolism and they are becoming ever so important in cancer biology.
Apart from the in vitro techniques involving FACS, TEM, metabolomics, enzyme assays and Seahorse what are some other ways to show mitochondrial dysfunction. More importantly, how do we show this in animal models.
SV40 is not considered an oncogenic virus in this review:
Article Oncogenic viruses and cancer
Why is SV40 not considered an oncogenic virus when it produces the large T antigen, which is used to immortalize mammalian cell lines?
Additionally, SV40 proteins have been found in human tumors.
Could this lack of coverage possibly have something to do with the fact that the polio vaccine introduced SV40 into the human population in the late 1950’s? How many people are actually positive for SV40 proteins? How many of these people develop cancer? Why are SV40 proteins not tested for regularly, given that they are an indicator of cancer?
AF2 may most strongly impact experimental determination of protein structure by multiple methods with possible benefits and negative impacts over time.
Some areas may need to quickly pivot or become obsolescent, whereas other may thrive.
Structures of large macromolecular machines should be enabled by having accurate computational structures for subunits and components.
The already great value of sequence data (which is doubling every 8 months) is likely to become far greater by being more directly connected to spatial information.
Overall the pace of biology and biophysical advances can be expected in increase by the ability to better harness the flood of sequence data.
What do you think?
Recently, I am curious to know about mechanisms of endometriosis and signaling pathways.
I have a question about the difference between signaling pathways in benign tumors and malignant tumors.
Since I studied, I noticed that the signaling pathway involved in benign and cancerous cells is similar, like MAPK signaling, Wnt Signaling, Apoptosis, Cell adhesion and angiogenesis.
So, what is the difference between endometriosis and ovary cancer in terms of pathway ?
Thank you in advance.
A study by Peeters et al. (2017) suggests that sugar traps cancer in a 'vicious cycle' which make it more aggressive and harder to treat (1). On the question-and-answer site Quora, Ray Schilling, MD, concludes: "there is a connection between the consumption of sugar and starchy foods and various cancers in man. Animal experiments are useful in suggesting these connections, but many clinical trials including the Women’s Health Initiative have shown that these findings are also true in humans. It is insulin resistance due to sugar and starch overconsumption that is causing cancer" (2).
1. Peeters K, Van Leemputte F, Fischer B, Bonini BM, Quezada H, Tsytlonok M, Haesen D, Vanthienen W, Bernardes N, Gonzalez-Blas CB, Janssens V, Tompa P, Versées W, Thevelein JM. Fructose-1,6-bisphosphate couples glycolytic flux to activation of Ras. Nat Commun 2017; 8: 922. doi: 10.1038/s41467-017-01019-z. https://www.nature.com/articles/s41467-017-01019-z.pdf
2. Schilling R. Why isn't sugar portrayed as bad like cigarettes? https://www.quora.com/Why-isnt-sugar-portrayed-as-bad-like-cigarettes
I am staining patient blood sample slides for identifying CTCs and I use the epithelial marker pan cytokeratin and mesenchymal marker Vimentin. I stain WBCs for CD45 marker. And I have seen that almost a big chunk (~60%) of my WBCs stain positive for vimentin . I am using the technique of immunofluorescence. Can someone please tell me the reason for this?
Guanine-rich DNA sequences can fold into four-stranded, noncanonical secondary structures called G-quadruplexes (G4s). G4s were initially considered a structural curiosity, but recent evidence suggests their involvement in key genome functions such as transcription, replication, genome stability, and epigenetic regulation, together with numerous connections to cancer biology.
The capacity for guanylic acid derivatives to self-aggregate was noted over a century ago. Some 50 years later, fibre diffraction revealed that guanylic acids form four-stranded, righthanded helices leading to a proposed model in which the strands are stabilised via Hoogsteen hydrogen-bonded guanines to form co-planar G-quartets.
However, why guanine specifically? Why not adenine, thymine, or cytosine quadruplex?
Does eosin stains collagen in IHCs in typical H&E staining? Please share your thoughts below as comments. Please add a reference if possible (much appreciated).
For example CD44 is a stem cell marker, whereas Vimentin is an EMT marker. Mesenchymal cells are called MSCs (Mesenchymal Stem Cells), then why are there different markers for stem cells and mesenchymal cells?
I have some questions related to a drug called Cobalt 2 chloride hexahydrate.
I am currently preparing an experiment related to Hypoxia. I bought a drug called " Cobalt 2 chloride hexahydrate" (from Sigma Aldrich) with the molecular weight of 237.9 g/mol and the total weight is 5g.
I want to know what is the best stock concentration should we make for this drug? Plus, how much H20 should we add to the drug ? and the optimal condition to store this stock concentration?
Furthermore, can you recommend me the best final concentration of this drug to mimic the Hypoxia environment? any ideas?
I have ordered KSFM media kit but the supplier forgot to send the BPE with the package. I need to run an experiment using NP69 as normal panel cells for anticancer drug test. I have FBS laying around, is it advisable to use FBS while waiting for the BPE delivery? The media KSFM has supplement growth factor and human recombinant EGF added.
Oncology is a branch of medicine that deals with the prevention, diagnosis, and treatment of cancer. A medical professional who practices oncology is an oncologist. The name's etymological origin is the Greek word ὄγκος (óngkos), meaning 1. "burden, volume, mass" and 2. "barb", and the Greek word λόγος (logos), meaning "study".
Cancer survival has improved due to three main components: improved prevention efforts to reduce exposure to risk factors (e.g., tobacco smoking and alcohol consumption), improved screening of several cancers (allowing for earlier diagnosis), and improvements in treatment.
I'm interested in the required knowledge,skills and branches of science that I may need to study in order to make a research in nano-medicine for cancer therapy. A list of references or something similar would be of great help to me. Thanks for all your kind responses.
Is it because we are trying to target these cells before they have a chance to divide??? I guess I don't understand the logic behind this.
I'm focusing on the Ras/Raf/MapK/Erk pathway and I'm expecting to see both when I treat with a MEK inhibitor in pancreatic cell cancer line. While I dramatically reduced levels of pERK, there's VERY little cell death. Does this mean that apoptotic pathways aren't necessarily rescued in this case or it is a technical issue where I should leave my cells in the treatment for longer than my current timepoint?
Our lab is currently testing the NTA (Nanoparticle Tracking Analysis) technique with the ZetaView from Particle Metrix. However we are really surprised how variable measurements are and how easy it is to get the results you are expecting just by changing the parameters randomly. It seems to have a lot of limitations. Nevertheless NTA is used widely in the EV field.
I tried to measure the fractions of an Optiprep density gradient to get an idea how many particles are present in each fraction. Is it possible to measure all fractions with the same settings and to compare the absolute particle numbers? Particle Metrix told us that different subpopulations might require different settings. That is why I was asking myself how I should adjust the settings to the different EV populations that I find in my gradient fractions.
Thank you very much for your help.
I have been imaging many z stacks of Drosophila mid/hindgut tumors using a leica confocal microscope. I'm wondering what the best way to quantify these tumors would be. They are GFP+.
I know both leica and image J have quantification programs. Any preference? Any recommendations would be greatly appreciated!
I am currently doing senesecence-related beta-gal staining. The problem is I have to spend a lot of time to count the cells, which was indeed countless.
Does any one have an idea about how many cell counts are considered "enough"? How many different scopes should be taken in consideration of repitition?
There are a lot of published data that showed association between vitD deficiency and many disorders e.g. diabetes, metabolic syndrome, CVD, cancer,...
I have two questions, please shed some light on this. I am new to in vivo studies and some may find my questions are trivial. TIA.
My first question is,
I wanted to develop the MCF7 mouse xenografts. I did a search and came to know that the injection of estradiol is recommended to develop tumours prior to injecting the MCF7 cells. Also, I have seen some papers without estradiol. Can you tell me the role of estradiol in tumour formation? Can the mice form tumour without estradiol?
Once the tumour reaches a definite volume, I wanted to inject my compound of interest. Here is the question, which is a simple way to inject? I have found the following ways related to my studies.
2. Microosmotic pump
4. Oral gavage
Please recommend me a simple way to introduce my compound to the mice. If you have a better option also welcome.
Many thanks in advance,
I want to know whether tumor cells share their information by time passing. I were wondering if anybody could answer my question and introduce some good resources in this regard?
I once had a colleague in a university, he was a professor of postgrad studies. About three years ago he suffered a bladder cancer, see for example: http://www.cancer.org/cancer/bladdercancer/. Then he took a surgery abroad, but it seemed that the cancer was spreading. So he decided to take herbal remedies besides taking chemotherapy.
I am not sure what happened then, except the fact that two years ago he passed away. I dont know exactly if his condition worsened because of cancer grew or not. But this story makes me ask about the safety and effectiveness of herbal remedies. Some people think that herbal remedies have better credibility over other alternative medicines.
So do you agree that herbal remedies are safe for cancer treatment? Do you have experience. Thank you.
For a background on herbal use for cancer, see for instance: http://www.cancerresearchuk.org/cancer-help/about-cancer/treatment/complementary-alternative/about/harm/the-safety-of-herbal-products-and-medicines
i am currently looking for an experiment alternative to southern blot. I heard that pcr based assays give results quickly ( let's say within few hours). I want to perform sybr green relied pcr to detect copy number of my interested gene in the human genome like in southern blot. however, I couldn't find a proper protocol. If you address a protocol or share your experience, I will appreciate.
Tumors in animal models, mostly mice, are usually very “young” and immature. We inject cancerous cells in matrigell and start administering nanoparticles within the next few days or a week. In the case of human, cancer grows sometimes for many years, so cancer vascular system is much more developed and matured. Can we then compare passive (EPR) targeting in mice and humans? Are there any indisputable published data which proves EPR (based on nano size only) effect in humans?
I need some good cancer biology book recommendations, preferably both dealing with fundamentals and the books that would help me for my masters research course in cancer biology.... I would like a book that deals with both basics and other advanced topics in cancer.. Thanks in Advance...
In this article fig. 1. The Author has made a decision tree with 24 nodes. Each node is specified for a specific cancer tissue of origin and the couple of MicroRNA which can identify these cancer tissues of origin. My question is, if I isolate miRNAs at the node14(hsa-miR-21, let-7e), node21(hsa-miR-205, 152), node24 (hsa-miR182, 34a, 148), node10(hsa-miR-194, 382, 210), will it be enough to identify cancerous tissue originated from the lung.
Why am I asking this question, because, I want to identify cancerous tissue, which has migrated to different region but originated in the lung. So if I take miRNAs from those specified nodes, will it be enough to identify lung cancer tissue, which has migrated to different region but originated in the lung.
I've been trying to isolate extracellular vesicles on an optiprep gradient in 5-ml tubes. I've tried 2 and 12 h centrifugation and in both cases, the fractions seemed to be mixed after centrifugation and the vesicles mostly remained on top of the gradient. I never have this problem when I use 38-ml tubes (SW32 Ti rotor) for 2 h but for a particular experiment I need to run this in small volume. What factors should I change in my protocol when using smaller centrifuge tubes?
The War on Cancer refers to the effort to find a cure for cancer by increased research to improve the understanding of cancer biology and the development of more effective cancer treatments, such as targeted drug therapies.. The signing of the National Cancer Act of 1971 by United States president Richard Nixon is generally viewed as the beginning of this effort, though it was not described as a "war" in the legislation itself.
War stated as “a state of usually open and declared armed hostile conflict between states or nations” in Merriam-Webster dictionary. War indicated attack of foreign army.
On the other hand meaning of rebel is opposing or taking arms against a government or ruler. Something has to cause rebellion. There are complaints. These complaints have usually been ignored over so long a period of time that people have become impatient with the slow pace of change; they begin to feel that conditions are unbearable. These complaints are most important causes of rebellion.
I think that cancer cells are rebels (cells live in bad conditions) and they try to live .
We must change our philosophy. We must accept cancer cells as rebels not enemy and we must try to correct bad environment to calm rebels.
I am using gastric cancer cell line for my research, i used Lipofectamine 2000 for transfection, could any one recommend me the good concentration of G418 and Puromycin?
I am working with a PDCL cancer cells and I want to transfect GFP in the cells. I am using lipofectamine 2000 and I incubated it with the cells for 6 hour and seems to be good but after 6 hours they start to show some death due to Lipofectamine toxicity.
So I only incubated cells with the Lipofactamine and the vectors for 6 hours and then changed the medium to the normal medium that I am using for the cells.
I used in this experiment 10.000 cells and they showed some positive cells after 72 hour however they all died when I tranfered them to a larger T25 Flask,
Any suggestions ?
I am using MDA-MB-231 and MCF7 for doing experiment. We have CO2 independent medium. Could you tell me how should I add other components (FBS, Glutamin/max, Penicillin, Insulin..). If I need 1ml medium, how should I dilute these components?
For FBS, Glutamin.. should I store in refrigerator (2-8 degree) for long time or I have to freeze them?
I look forward to hearing from you.
Thank you so much for your help.
For example, breast cancer cells in mice models. Can transfected cells be used (assuming the transfection efficiency is high)?
I can not use the estrodial pellet, because of the price, so I have a plan to maeke MCF-7 xenograft model w/o estrodial pellet.
If I inject directly or O/A 17beta-estrodial in DMSO to mice, will that stimulate MCF-7 growth in mice? Or is there some other method to make estrodial pellet?
Genes transient transfected into cells will only be stable for several days. However, xenograft tumors formation need more time about 3 weeks. Whether or not we chould use transient transfected cells for xenograft tumors
We have created MDA-MB-231 stable knockdowns vs controls for our gene of interest. To finish off the in vitro data and publish our work we wish to use a xenograft mouse model to look at metastasis and tumor growth differences etc in vivo.
Does anyone know a service provider that can do this work to complete our project?
Most services appear to use xenografts for drug testing using their own in house cell lines, where as we wish them to test our knockdown and control cell lines.
I want to inoculate MCF7 or T47D cell lines after female albino mice irradiation. I read that it requires E2 pellet and that cell lines should be injected with Matrigel. Could you send me your recommendations about these points and the time needed for tumor to appear for these cell lines (both subcutaneous and orthotopic)?
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?
Recently we tried to transfect a plasmid into MDA-MB-231 and MCF-7 cell lines. After transfection we found that its transfection efficiency is different from other cell lines. MDA-MB-231 cell line is difficult to transfect with lipofectamin2000, after optimized transfection condition we only reached about 1% transfection efficiency. Has anyone performed transfection to this cell line with lipofectamin2000? Can anyone offer suggestions?