Questions related to Cancer Research
So I'm new to cell culturing research and synthetic biology and I'm working with MDA-MB-231 cells. Fairly regularly I have to at least split plates in order to maintain the cells, but I am also passaging cells for other things. Normally on a 10cm petri dish I'd plan to seed ~5.0E5 cells/mL for regular splitting and in a 96 well plate I'd seed ~4.5E4 cells/mL for my experiments. One thing I'm confused on still is using the hemocytometer to calculate cell density because I'm not entirely sure what my dilution factor should be.
Normally, I'll remove the old media, wash with PBS then trypsinize cells, after incubating for 5mins I add a bit more media until the total volume is 10mL, then I spin everything down. After that, I aspirate out the supernatant and am left with a small pellet of cells that I then break up and dilute with ~2-6mL of fresh media before using the hemocytometer to count the number of cells in 10uL.
My question is since I'm removing all the old media after centrifugation but then am adding new (different amount) media prior to counting, how does that impact the overall dilution factor?
Here's an example of what I was taught to do to find the amount of cells in 1mL, I just wanted to confirm that I'm on the right track. I was told if I resuspended my pellet in 3mL of media and then counted 75 cells I'd have:
75 cells/10E-4mL media
75 E4 cells/1mL media
7.5E5 cells/1mL media --> multiply this by 3 since you used 3mL to dilute before counting to get total number of cells (is this what accounts for the dilution factor?)
Total of 2.25E-6 cells that could be split however you see fit? Sometimes determining how many plates to get out can feel like a guessing game. Let me know if you see any issues with my method or suggestions that might be helpful, I welcome them. Please do try to be kind as I'm still learning and figuring things out. Thank you.
I am analysing several drugs on tumour cell lines. I need a quick way to calculate IC50 based on data generated.
In case of mTOR I notice cancer research wants to block this pathway right? So would resistance training, amino acids and insuline be dangerous to some extent? Someone with genes for cancer growth can he or she also have more danger with training?
Or is it only when high doses of insulin, growth hormon and such are ingested or injected?
Could you please provide any tips or ideas you have for creating the finest meta analysis review on cancer? Rather than writing a review article, I'd like to begin working on a meta analysis.
We want to know if one protein can be used as a breast cancer biomarker or not. So we'll have to compare normal and patients, and our question is whether we can get samples from patients who received chemotherapy before?
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.
in cancer therapy, sometimes you have to target multi-targets to understand the molecular pathway of the specific protein. CHyMErA can target multi targets but it is not safe and has a high risk of extra mutation!
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!
I'm trying to design a simple cube-shaped DNA origami with specific dimensions from just single stranded DNA oligos.
Since there is no specific scaffold the origami is based on, cadnano is difficult to use...
I understand you can use Nupack... but I'm not sure how to create 3d objects using Nupack.
Would there be a better program fit for this or what would be the ways doing so in nupack?
I want DNA microarray cancer data sets with features and samples to form a gene expression matrix for applying intelligence techniques (such as basiayan neural network algorithm) to classify cancer data sets. Please help me.
Permissible limit of arsenic in drinking water was 50 μg/l by WHO which was later reduced to 10 μg/l. Currently many countries like Bangladesh and China have not updated their own Permissible limits. However, this is been observed that on the event of long term exposure of arsenic at low concentration that is less then 10 μg/l in drinking water, health complication arises.
I made a stock solution (25mM) of Pioglitazone hydrochloride (Tocris) in DMSO. It has dissolved finely. Then I treated the cells maintained in serum free DMEM with a final concentration of 100 micromoler of Pioglitazone. Here it forms crystals, precipitates in the culture vessel, but I am not getting the desired effect of it on the cells. Please suggest how to achieve the proper effect of Pioglitazone treatment?
There is an increasing amount of research on the ketogenic diet in cancer therapy. However, I can't find much about this diet in those guidelines produced by governmental cancer research institutes such as American Institute for Cancer Research (AICR). As a matter of fact it seems like the word ketogenic is totally missing from the databases.
Am I missing something?
National Library of Medicine results: https://pubmed.ncbi.nlm.nih.gov/?term=ketogenic+diets+cancer
The main ‘party trick’ of cancer stem cell (CSC) seem to be that it provide another explanation for tumour heterogeneity (i.e. cell divide asymmetrically give rise to progeny cells that are different). Secondly, asymmetric cell division is what makes this model ‘useful’ (i.e. clinically relevant): CSC model refer to a subpopulation of cells that must be targeted for effective thematic outcome. But if asymmetric cell division does not take place (i.e. if all cells divide symmetrically) than any therapy that kill cancer is in fact targeting the CSC (i.e. there is no point in identifying CSC- each cell would be one under symmetric cell division).
So, is asymmetric cell division a hallmark of CSC model? Any good literature on this?
I am trying to find a good immunocompetent mouse model to study triple negative breast cancer (TNBC). The 4T1-BALB/c model is perhaps not ideal for studies on immunosuppression and as 4T1 is derived from BALB/c mice, it cannot be transplanted in the C57BL/6 strain. Can someone suggest a good animal model to study TNBC-induced immunosuppression?
I have been injecting AML primary cells in NGS-SGM3 mice and they developed leukemia. After recovering the human AML leukemic cells from the bone marrow it is difficult for me to keep them in culture and especially to expand them. I am using SFEM media with CC100 cytokine cocktails + Tpo. This media worked well for primary AML but is not giving me good results with the same cells isolated from the mice/xenograft models.
How can I optimize the condition to maintain AML cells recovered from the mice?
I had a contaminated cell culture I suspected several things, including the media. I put the media (only) in a T-25 flask and used 2 agar plates to check bacterial contamination (one opened in the laminar air flow the other in the CO2 incubator) and left them is in the CO2 incubator at 37 oC (after I cleaned it),
two days later I didn't find any thing on the agar plates and no changes in color or clarity of the media in flask however I examined the flask under the microscope and found these things .. What could it be?
I am working with rectal swab samples in the field of colorectal cancer research. Does anyone have experience (or can point towards any relevant literature) of specifically working with proteins (extraction, characterization, or anything even remotely connected to protein work) in rectal swab samples (or any mucosal swab samples).
I just asking cancer expertise to suggestions the best books can read by MSc students before start their PhD program in cancer research
I'm fairly new to cancer research and there's a host of articles on Sunitinib being administered via oral gavage in mice models but I'm unable to find any information on studies that have explored directly injecting tumors with Sunitinib. Just curious.
I've been writing a review article (almost done) on cancer research. Is anyone interested in collaborative writing and publication? Please let me know. We can share our ideas on extending the content of the paper and get it published mutually.
Professionals with an interest in cancer research are most welcome.
I currently doing a research for anticancer drugs and have a plan to use CA 15-3 as tumor marker to measure the effectivity. Some reference tell that CA 15-3 and MUC 1 are synonym, but i also found some reference that stated CA 15-3 and MUC are different. My question is "Could I use ELISA kit for MUC 1 to measure the CA 15-3?" because my local supplier can't find the ELISA kit for CA 15-3 in affordable price.
I read many papers about exosome isolation from fresh blood or frozen plasma, but not for frozen whole blood. I am wondering whether can I get exosomes from cryopreserved whole blood? One of my workmate told me the blood cells were broken if crypreserved and would affect the isolation of exosomes? What I need is the exosomes and their surface biomarkers. Could anyone give me your prescious advices? thank you so much.
Could anybody please tell me what are the different types of cells that originate from HNSCC patient samples. What are the type of studies that it can be used for?
I understand there are epithelial, fibroblasts, and keratinocytes that can be derived from HNSCC tumor samples. Can immunological studies be done using these type of cells?
I isolated a large amount (I hope) of exosomes and I need to quantify the membrane proteins with a Bradford assay before doing a WB. I started off with 70 ml of conditioned medium which I managed to concentrate to about 1 ml before ultracentrifugation. After ultracentrifugation I added 200 ul PBS to my samples. My expectation is that the concentration of exosomes is quite high. So now I need to do a Bradford assay and I would like to know how to dilute my samples.
I am new to cancer research and I am trying to optimize my small cell lung cancer cell protocol.
I purchased a cell line from ATCC (H1105) and I have been growing the cells in RPMI 1640 media (10% FBS + penicillin/streptomycin.
I am looking for anyone who was worked with these cells in the past and who can let me know if these cells look normal?
I noticed the cells grow very slowly and form spheres.
Am I using the right media? How can I get the cells to grow faster? Can someone refer me to a good SCLC cell culture protocol? Thank you.
I need to confirm a knockdown of HER2 in SKBR3 and BT474 cells, have had problems blotting for HER2 in the past so any advice with regards to the protocol or antobody choice is much appreciated, thanks
I have transfected GFP-AR in to HEK293 and LncAP cell lines in serum free medium (HEK293-DMEM and LncAp-RPMI1640) using Lipofectamine. After how long should i change the medium? and should change it with serum free medium or serum containing medium (10% FBS)?
I am studying on cancer research and ı used bcl 2 antibody to understand there is apoptosis or not. I got good band ı tested with ponceu red. Finally I got this result.ı used uvp chemi doc it2 imager to preview membrane.I tried a lot of times what do you think about this result?
Tumor suppressor genes (such as Rb, APC, PTEN and the ever famous p53) are often downregulated in cancer cells, if not completely lost or dysfunctional.
I'd like to know if there are any tumor suppressor genes that are upregulated in metastatic cancer cell lines/models? If so, could you link to articles/publications/studies of such genes? Are these genes bi-modal? Or have people figured out the molecular mechanism behind these genes?
I'd appreciate any and all help provided.
I am planning to use the Qiagen RT2 Profiler PCR array for signaling pathways. Is it really necessary to use the RT2 SYBR Green Mastermixes and the RT2 First Strand Kit with the profiler array? Can I use Applied Biosystems SYBR Green PCR mastermix instead (Cat no. 4309155) as well as my already synthesized cDNAs? The platform is Roche Light Cycler 480.
Looking forward for some help. Many thanks.
Dear friends and colleagues,
there is a very important resource on the web that is "Atlas of Genetics and Cytogenetics in Oncology and Haematology". The Atlas is a peer reviewed on-line journal / encyclopedia / database established in 1997 that collects data about cancer genes, solid tumors, leukaemia and other resources. It is in open free access for readers and there are no fees for the authors.
I'm searching volunteers and collaborators that would to write with me reviews about single genes involved in cancer.
I have published several papers so far and I believe that Atlas is a very valuable tool for us researchers, scientists and scholars.
I invite you to visit the Atlas website at http://atlasgeneticsoncology.org/index.html for more information. Here is my latest article published http://atlasgeneticsoncology.org//Genes/GC_EEF1B2.html to show you my working method. Anyone wishing to collaborate with me to write a specific review on a gene can tell me as well.
recently I am working on a project about the effect of bio-luminescence in cancer research. It would be my honor and pleasure to help me on this way
The event of death by metastasis or recurrence is very common, many researchers have linked the tendency of some tumors to re-appear to the presence of occult or dormant cancer cells that may have a phenotype that allows them to remain "hidden" from the immune response. However, the understanding of these cells and the mechanisms that they use to achieve evasion remain mostly unknown. It is critical to understand these cells better and to be able to detect their presence for they are of great therapeutic importance.
There are a lot of published data that showed association between vitD deficiency and many disorders e.g. diabetes, metabolic syndrome, CVD, cancer,...
I am looking for information concerning the effect of publications regarding 5-year survival of diseases, like cancer, on the feelings of the patient. As the statistics are frequently published, and available on many websites with information on serious diseases, I was wondering whether psychological research was done as to the impact these statistics have on patients, dealing with diseases. I have tried to find such research myself, but the overwhelming majority of research I have found deals solely with diagnostic importance of these analysis. I will be most thankful for any advice and references on this issue.
Next week I have my first 'serious' interview for a postdoc position job that REALLY interests me. It would be done on Skype, I was asked to prepare 20-minutes powerpoint presentation on my 'previous scientific work'. Since I don't have experience, I am not really sure what does it mean. Should I make a 'classic' ppt with structure: introduction, aim, materials&methods, results, conslusions of all my projects? (Would be difficult in 20mins) Or should it be more like what was the project, why we wanted to study just this, what techniques I used, what I learned, what are my skills for the moment etc?
I will appreciate any advice :)
PS: we are talking about cancer research position
several old studies says no, more recent studies says yes. Robert Bell in 1870 used raw salad to increase amygdalin in cancer patients and used saliva to detect the presence. Several sources say amygdaline converts to cyanide in an acidic enviroment and when gets in contact with acid lactic of cancer masses convert to cyanide and kill them (sort of selective killing). To be absorbed as amygdaline and avoid convertion into cyanide in the gut it should be taken from food rather administered directly. There are some roman texts about incresing lifespan of cancer patients with regular intake of 2-3 bitter almonds (rich on amygdaline) a day. Anyone has experience on it? Are research on amygdaline extensive and satisfactory? What is your opinion?
Can anyone suggest me the precise and new anti-cancer target for docking of peptide type compounds. As per literature reports the chemical class of the above peptides are mitotic inhibitors. The BAX and BID along with Caspase 3 were highly expressed with peptides while in cytotoxicity studies. When the above genes are over expressing (Biomarkers) is an indication of apoptosis. Can I choose Caspase 3 as druggable target for these compounds and go for docking ?
Cancer research has developed precise biomarkers to use a proper cure for different kind of cancers. Why there is no proper biomarker to find the precise treatment in autoimmune desease, such as psoriatic arthritis?
Since humanized and SCID mice are very expensive to procure and maintain. I was wondering to develop a cost and effective approach to develop an in vivo model for cancer research. Thymectomy is well established method to cause immunosuppression. I was wondering what will happen if we try to develop PDX model in thymectomized alone or in combination with immunosuppressants in C57bl/6 or Balb/c mice. Will tumor model be developed ?
I have just started using Olympus fluoview for confocal microscopy and I don't know much about microscopy. I plate my cells (HeLa) in 4 welled Chambers slide. Stain the cells with Mito tracker dye, fix it with 4% PFA, wash, add mounting media and do the microscopy. The first time everything was fine. The second time I was trying to optimize the dye concentration, but something went wrong. The microscope kept on showing 'The focus position not found in scanned containers'. I had two slides and both of them had the same problem. When I tried with my old slide, it was fine. I have no clue what is wrong with my slide. All the chemicals and reagents that I have used is same as before except for PFA. The second time I diluted 8% PFA with sodium phosphate buffer to get 4% PFA. Can PFA cause problem finding focus?
We are trying to test some formulations of PD1 antibody on cancer models and would like to have a robust in vitro technique for future in vivo assessments.
Most studies on CAR-T cells use NSG mice (B/T cell deficient) based on the papers I found. Only a few use nude mice (T cell deficient, B cell present), which is what we are hoping to do. Does anyone have experience with using nude mice tumor model in CAR-T cell studies? What are the problems with using nude mice for CAR-T cell studies (since B cell is APC), and what are the potential solutions? Thanks a lot for any responses!
In my job, I transient transfected Active-wnt3a construct into Hek293T and Hek293 for expression , I collected three batch(fresh medium 5ml in 10cm dish for 24hr)
conditioned medium and concentrated in 500 µl ,but it didnt have any effect when it treatment 20µl for TopFlash repoter assay!!!!
Can any one tell me what can i do?
I have submitted a manuscript to clinical cancer research and the status changed to "decision pending" on the same day "under review". And I wonder whether it is not good news, how long will I receive the email from the editor?
We are looking for highly motivated candidates to join our lab. More info available in the link below:
Any help with spreading the word about them will be highly appreciated.
Hello! For my study assignment (cancer research, up/downregulation of a gene in high and low density cells) the standard deviation is not necessary but we think it is important to put in our short communication. We found out that the standard deviation is taken from the delta-CT value. For example: you have a gene of interest and a reference gene. Each gene is done in triplo and you have a control and a treatment.
GOI control: 32.37, 31.76, 31.80
Ref control: 16.05, 15.10, 15.76
GOI treatment: 30.1, 29.78, 29.88
Ref control: 18.56, 15.64, 14.76
We've calculated the delta-CT from the averages of the measured values. How can I calculate the standard deviation for the delta-CT value?
Thanks in advance for your help.
I'm a Research associate at 57357 hospital , Egypt , I would like to ask about two cell lines : RD , RH30 ( Rahbdomyosarcoma ) we couldn't find them in Egypt and we are committed to certain budget , deadlines and masters thesis based on this project , anyone know a trusted source or a lab in any country usually uses them to give as aliquot ? Thank you in advance !
What could we change in preparing tissue samples prior cryosections(cryostat) method for more convenience and results? ( I mean cryo-embedding) The second question is in which areas of research do you use this one and why is it a pretty rare method of imaging? It is interesting to get answers from researchers who are working currently with this method or those who worked with that recently.
Thank All in advance for any ideas.
I'm starting to work on microRNA profile in a mouse model of liver cancer. We would like to use the next generation of sequencing approaches since it enables mapping and quantification of all transcripts.
Since it is the first time we are working with sequencing, I was wondering if somebody could suggest the best company to contact. I was in contact with LC Science because a colleague of mine recommended them. We are also evaluating the Arraystar sequencing service. Does anyone have some suggestions or recommendations for other companies?
I plan to perform CAncer Personalized Profiling by deep Sequencing (CAPP-Seq) for a cancer research project.
Is there any available genome analysis company that uses Illumina Hiseq or a similar NGS system and that can perform CAPP-Seq with a reasonable price?
We will need logistic support for cargo shipment from İzmir in Turkey as well.
I'm looking to find a way to determine the concentration of the ketone body Beta-hydroxybutyrate (BHB) within cell culture media. Preferably, a method that would employ a spectrophotometer. Is it possible to make up a solution of known concentration and run it through a variety of wavelengths in order to determine the lambda max for the ketone body? Once I have the lambda max, shouldn't I be able to determine the concentration of BHB within a solution by performing a serial dilution of BHB solutions of known concentration, then comparing the unknown solution's absorption to those of the standards?
If anyone has a better way to determine the concentration of BHB within a solution, then I would love to hear it.
Thanks in advance!
Transhiatal esophagectomy was championed by American Dr. Orringer since 1976. The issues were: inadequate field exposure; inadequacy of node clearance, use of blind & blunt dissection & (rarely) azygos vein tear. Benefits: avoiding thoracotomy and all the morbidity associated with it.
So, what is the current opinion?
I was wondering if anybody have used stain free gel (Biorad) to compare the expression of a protein of interest in Cytoplasm and Nucleus?
If I were to submit a paper including this stain free blot image to a decent journal, say Cancer Research, would that be any issue?
I am wondering whether I have to go for conventional normalization method(using loading control) for comparing proteins from different compartments in cells. While stain-free blot is easier to work with, I am worrying that reviewer would still like to see loading controls of nucleus and cytoplasm.
Hello fellow Patchers and Sealers,
I made a pipette solution recently and expected the osmolarity to be around 270 mOsm approximately according to rough calculation but when I measured it it was only about 191 mOsm. I am pretty confident I did not miss out on any ingredients or added more water that could decrease the osmolarity than expected. Now, at this point, I am requestioning my manual calculation of 270 mOsm (approx) from the recipe. It would be really helpful if someone can proof-check my calculation for the recipe below. Just the K- gluconate itself should make it 244 mOsm to begin with.
NOTE: To adjust the pH to 7.4 I had to add about 1 mL of 1M KOH. I wonder if this could be a reason. I doubt it though since only an mL of this in a 100 ML final volume should not make the difference.
Recipe: K-Gluconate 122mM, NaCl 9mM, MgCl2 1.8 mM, Mg-ATP 4mM, Na-GTP 0.3 mM, Creatine phosphate 14mM, EGTA 0.45 mM, CaCl2 0.1 mM, HEPES 10 mM