Pancreatic Cancer - Science topic

I read that Israel has developed a cure for pancreatic cancer (see link below) Can anyone tell me more?

Hello everyone,

I am cultivating patient derived organoids from pancreatic cancer patients and I saw at the bottom of my domes some "strange" cells other than my usual organoids. I know I should perform some IHC to understand the identity of these cells but I was wondering if some of you could help me to have a general idea of their possible origin. They don't seem fibroblasts to me, so maybe are they normal ductal pancreati cells?

Thanks in advance

I am working on pancreatic acinar cell function.

I have checked so many papers it seems there is no acinar cell lines or organoids so far.

I am wondering like-minded researchers who work on acinar function can have other alternative methods on this topic, especially in vitro research!

Thank you so much!!!

I am planning to do gene knockout for human cell line using all in nonviral plasmid which has both the GFP and the Puromycin resistance gene. I am wondering which one is better to start with, the cell sorting with GFP and then The Puromycin selection, or the opposite?

CRISPR/Cas

#Pancreatic Cancer

#Biotechnological Engineering

#Biotechnology

#Molecular Cloning

#Gene Knockout

#Genetic Engineering

#CRISPR/CAS9

#CRISPR

Hello everyone,

I am currently working on patient derived organoids from pancreatic cancer (PDAC) patients. I noted that my organoids never grow too much. after 1 month in culture they are usually around 100-150 um. I am following this very famous protocol ( ).

Do you have any suggestions?

Thank you a lot in advance

Giulia

I am receiving established orgnaoids from a different lab who use Wnt3a conditioned media in their organoid culture. I will be using recombinant Wnt3a (5036-WN-010/CF, R&D) and am wondering what concentration to use. I have seen 4ng/mL or 30-100ng/mL. Is there a standard for pancreatic cancer organoids?

I am working on pancreatic cancer.

Now, I am trying to generate a pancreatic exocrine organoids using my KPC mice.

But after I checking the protocol, PancreaCult™ Pancreatic Organoid Growth Medium (Mouse) | STEMCELL Technologies

I am not sure the component of the organoids, just containing duct epithelial cell? If there are some acinar cells in the organoids?

Thank you so much!

Previously I did some surface marker expression (CD40, CD86) of RAW macrophages and DC2.4 cells using flow cytometry. For RAW cells, results were not satisfactory. Now, I am optimizing apoptosis assay with a pancreatic cancer cell.

We are interested in CD8+T lymphocytes form pancreas cancer patients. Is it possible to find a pancreatic cancer cell line with HLA class I molecules that could be recognised by CD8+T lymphocytes from an individual patient?

I have been trying to culture cells pancreatic cancer cells and normal cells (GM05757). fIRST CELLS DONOT ATTACH TO THE FLASK EVEN FEW CELL ATTACHED TO THE FLASK BUT THEY DONOT DIVIDE AND AFTER 2,3 passages there is fungal infection.

I'm using panc02 cells, mouse pancreatic cancer cell line.Beforehand, I had some issues without doing any fixation, so now I’m trying with fixed cells.

Currently, I am looking for a solution to develop chemotherapeutic drug resistance in a primary cancer cell line. But after a quick look at the literature, it is indicated that the management of acquirement of drug resistance takes plenty of time, more than eight months! Is there any convenient method to subculture chemoresistant cell lines in a short time? Or any other suggestions rather than eight months interval with an increasing dose of chemotherapeutic agent?

Best regards.

Can anyone tell me how to stimulate pancreatic cancer cells for cytokine release in cell culture ?

Due to the rising burden of pancreatic cancer and poor outcomes, a precise,

post-operative cancer staging for further and individualized therapy is needed. In the latest cancer classification system, the lymph node invasion mechanism is not addressed. Due to different outcomes regarding the lymph node invasion, we suggest a rethinking of the current system.

What is your idea? Is there a difference between direct lymph node invasion (per continuitatem) and lymph node metastases distant to the tumor? Shouldn´t this be taken into account?

Some diseases can have specific membrane proteins as targeting markers that endow biomimetic nanoparticles with natural tropism to affected areas, such as CD138 in multiple myeloma. Which examples are there for Pancreatic Cancer?

I am updating my earlier review on the role of Fluoride doped Hydroxyapatite in Cancer and my current focus is on Psammoma Bodies which have been found, and identifed as high risk, in a very wide range of Cancers. These include Cancers of the Bone, Spine, Brain, Choroid Plexus, Dura Mater, Gliofibroma, Medulloblastoma, Meningioma, Cervix and Endometrium, Ovary, Kidney, Lung, Mesothelioma, Pancreas, Skin, Hemangioendothelioma, Olfactory Neuroblastoma, Duodenal Somatostatinoma, Stomach and Thyroid. Early studies did not have the benefit of advanced analytical techniques, or did not even consider the Fluoride content or composition of the mineralization. Can anyone help by supplying analytical data based on Raman spectroscopy, neutron activation, x-ray or wet analysis?

Could you please give me some advice? I wonder if I have a 40-mm3 pancreatic cancer tumor what the volume is in humans. In other words, does conversion of mouse tumors to human tumors exist? And what about the conversion considering the age of mice and humans? Thank you a lot for any ideas.

I'm preparing pancreatic cancer cells spheroids. Which concentration of collagenase can be used to dissociate them?

Kindly provide 1-2 papers showing evidance about the mortality rate in hamster model for pancreatic cancer development (using BOP).

We're just starting to isolate pancreatic fibroblasts from healthy and a tumorigenic pancreas but it does not work very well. We tried freshly isolated and also frozen tissue and use a collagenase IV digestion. Additionally we add FGF to the media.

post #for #concern #people #oncology #research #area

For metastatic pancreatic cancer apart from chemotherapy is there any other medical treatment (newly adopted) exists in India?

As you know in this stage the theoretical prognosis is very poor. Is there any other treatment by which we can treat the patient with best comfort?

CA19-9 is a commonly performed tumour marker in pancreatico -biliary malignancy,but the levels keeps varying ? depending on the severity of obstruction causing cholangitis.In these circumstances if the levels are high does it mean it is a disseminated malignancy or it is probably due to cholangitis !

I have a 96-well angiogenesis assay plate. For the first time, I will design an in vitro angiogenesis assay.

I'm going to test the potential for tube formation after treating the pancreatic cancer cells with a few drugs. I thought that pancreatic cancer cells would be seed directly into the wells and the potential for tube formation could be assessed. However, I have seen that HUVEC cells are used in the studies. Do I need to use HUVEC cells in this experiment? If yes, what is the reason for using it? Can you also tell us the tricks of the angiogenesis experiment?

Thanks for your help.

Hello everybody!

I've run a cox proportional hazards model for survival analysis in a cohort of pancreatic cancer patients with SPSS v. 25 and I want to

1. compare the accuracy (with the Harrell's C) of my model with classical staging

2. to measure the Cs of my model after bootstrapping

I've tried the macro available in the IBM web site, but it does not work (many errors)

...anyone can help?

THANKS

Is the use of HIPEC with resectable pancreatic cancer will affect the prognosis

Intraductal papillary mucinous neoplasms (IPMNs) of the pancreas are being diagnosed with increasing frequency. IPMNs comprise a histologic group that ranges from adenoma to invasive carcinoma with different degrees of aggressiveness (borderline tumor). Actually, it is not known whether all IPMNs have this malignant potential or what is the best treatment of IPMNs. The'identify the right time of surgical treatment or follow-up is the great dilemma .

Recently I started to do IHC experiments. I got some IHC stained slides, but I couldn't distinguish pancreatic cancer cells from normal tissue cells and other stromal cells. I found some references, but the images in these papers are too little for me to read the slides. Maybe I should find some books to read, but I don't know which book is the best choice for me.

I have downloaded the miRNA-seq data for pancreatic cancer from TCGA. Now, I would like to see the expression of a particular miRNA across all the samples - or at least to classify samples into two groups: high and low expression. What exactly should I do?

Soft pancreatic remanent.

Hello everyone! I have started a primary cell culture from murine heart, bladder and tight. I need to do a proliferation assay using EdU-Click kit, and I was wondering how long should I incubate the cells with the EdU. I optimized the protocol on AsPc-1 (pancreatic cancer) and PS-1 ( satellite cells) cell lines, and incubation of 1h was fine, as the cells replicate quite fast. But with primary cells I guess the situation is a bit different, they replicate slower, so I guess 1h is not enough. Has someone used Edu kit with primary cells? Do you have any suggestions?

Thank you in advance!

I am a beginner and having a hard time finding statistics reference for obesity/pancreatic cancer.

please help

Thank you

For finding a mutated cell line in ERRFI (MIG6), I use CCLE and COSMIC databases as well.

Both of them show that MIA PaCA2 has a Nonsense mutation in MIG6 and the protein length is 323 instead of 462 (wild type), but surprisingly after running the western blot for MIG6, I've found that the molecular weight for this guy is same with wild type cell lines in MIG-6 (for example Panc1).

Hello,

i'm writing a grant proposal for a study involving circulating tumor cells (CTCs) dynamic in pancreatic cancer.

I'd like to know more about devices for CTCs detection available on the market, about the main differences among their features and their approximate price.

The primary outcome would be just a total CTCs count. Discriminating between subtypes (ie epithelial, mesenchimal) would be nice but not mandatory.

What device has the best quality : price ratio?

Thanks!

Hi,

I am having some difficulty finding papers which used nanoparticles to treat pancreatic cancer using 3D pancreatic cancer cells. please is anyone able to recommend some papers to me?

Hi,

I am working on Gemcitabine resistance in pancreatic cancer cells (ASPC1, MiaPaCa2, HPAF2, CaPaN2 and BXPC3), and I should use the IC50 of Gemcitabine in all the experiments. I found different values in literature and the numbers are highly variable ranging from lower nanomolar to lower micromolar for the same cell line under same conditions!

The IC50 I got in my hand is also different from literature but its consistent as I got nearly the same value in 3 different experiments.

Should I use the IC50 I got in my hand even if its different from literature?

Thanks

Too many people die from late stage pancreatic cancer. It is diagnosed too late

Is there a good screening test that can be applied to people from the age of 50 and above?

I have to analyse the necrotic/apoptotic cell population for pancreatic cancer cells after treatment with some synthesized nanoparticles. The problem I am facing is that I do not have access to flourescent or confocal microscopy and I have to travel 3 hours to get the imaging done. How exactly can that be done without damaging the cells in the transit as the cells are not fixed for the following protocol. If I fix the cells then will AO and Etbr penetrate the fixed cells (fixed at the stage when they were damaged). Please provide the reference for the same.

Pak 1 expression in pancreatic cancer.

We have known that many advanced cancer patients are refused to treat more after standard chemotherapy at Shizuoka Cancer Center, even if they could be estimated to recover with other treatments including thalidomide and celecoxib. They are ordered to select palliative care even though they have a chance to be saved with other treatments.

Please help Japanese advanced cancer patients save their lives from Shizuoka Concentration Cancer Center (President Ken Yamaguchi). They have to die with the cease of respiration after injections of overdose morphine hearing the stream sound of high volume oxygen.

I synthesized highly purified thalidomide in 1999 and treated the patients since 2000. But we are ordered to recall every capsule from the Ministry of Health, Labor and Welfare in 2002. The reason is thalidomide is dangerous medicine. In 2005, we submitted a clinical trial, using thalidomide, celecoxib and low dose gemcitabine for pancreatic cancer, to the Ministry of Health, Labor and Welfare. But the clinical trial for multiple myeloma of Fujimoto Pharmaceutical Corporation with poor medical knowledge was approved by illegal transaction between bureaucrats and the Corporation. Fujimoto Pharmaceutical Corporation should conduct various efficient clinical trials to help the patients with advanced and metastatic cancers.

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 transfected BxPC3 cells with 5µg Akt plasmid using Lipofectamine 2000. I also had a control where I added only Lipofectamine 2000. Surprisingly, I observed 60-70% cell death in the wells transfected with Akt plasmid whereas no cell death was observed in the wells treated with lipofectamine 2000 alone. Below is the protocol I followed for transfection:

1. Seed 0.2 x 10 6 cells per well in a 6 well plate

2. Incubate overnight

3. Aspirate the media and wash with 1X PBS

4. Add 1.7ml of OPTI-MEM reduced serum media

5. Incubate at 37 o c for 45 mins

6. In the meantime, quantitate the plasmid and the plasmid concentration was 1182.2 ng/µl

I had 3 wells to transfect with 5µg Akt plamid and to avoid the losses I took lipofectamine 2000 and plasmid for 4 wellss

7. Mix 600µl OPTI-MEM + 36µl lipofectamine 2000

8. Mix 600µl OTI-MEM + 20µg Akt plasmid.

9. Incubate for 5 minutes at room temp.

10. Add 600µl of diluted DNA to 600µl LPF

11. Incubate for 8 mins at room temp

12. Add 300µl DBA-lipid complexes drop by drop per well

13. Incubate at 37 o C for 8 hours

14. Replace the OPTI-MEM reduced serum free media with fresh RPMI media with 10% FBS

15. Incuabte at 37 o C for 48 hours

Hence, each well contains 9µl Lipofectamine 2000 and 5µg plasmid

and I treated one well only with 9µl Lipofectamine 200 where no cell death was observed.

Any suggestions would be of a great help to me. Please assist in this situation.

I am developing a cancer mouse model. I injected a pancreatic cancer cell with two variant WT and KO(some gene) in nude mice subcutaneously. After 4-5 days all mice died in KO batch but all mice are still alive in WT group. What could be the possible explanation for that?

I am trying to transfect GFP labelled plasmid to pancreatic cancer cell line. (miapaca-2, panc-1) with 2ug dna and 8 ul lipofectamine LTX with 2 ul plus. in serum free media and then left it for overnight and add complete media after 18 hours. but i got only 30-40 % transfection efficiency, is their is any way to increase this efficiency.

Could anyone provide me some suggestions which xenograft model to use for evaluation of anticancer activity of a therapeutic agent - ectopic or orthotopic? I am involved in in vivo studies related to pancreatic cancer. For your information, I have read about these two models and have some idea on the pros and cons of both models. However, I could not make a decision on which xenograft model to use for my studies. I will use Ncr nude mice and the pancreatic cell line Capan-2. 

Kindly assist me with this question. Thank you.

I could not found the cell sizes of some cancer types such as; lung (NCI-H2126, ATCC CCL-256), breast (UACC-1179, ATCC CRL-3127), over (EFO-27, DSMZ ACC-191), melanoma (A-375, ATCC CRL-1619), pancreatic cancers (PA-TU-8988S, DSMZ ACC-204).

Could you share your information?

Thank you for your help.

I am growing pancreatic cancer organoids from patient biopsies on matrigel using a culture media for pancreatic organoid growth. My cells usually form organoids nicely, but some are eventually overrun by fibroblasts even after passaging. Does anyone have any suggestions for eliminating fribroblasts from my culture without killing the organoids?

The data set should include patient demographics, symptoms, family medical history, comorbidities, certain lab test results etc.

Thanks

A recent meta-analysis has shown that pancreatic elastase test has a combined sensitivity of 0.77 in detecting exocrine pancreatic insufficiency (Vanga, Rohini R. et al. Clinical Gastroenterology and Hepatology, 2018). What's the impact of this reported sensitivity in current clinical practice? Would a test with higher sensitivity make a positive and significant impact on time-to-diagnosis?

I am interested to test the efficacy of my compound on pancreatic cancer cell lines. NCCS pune dont have them. Can someone share their freeze or help me to get one. (  except ATCC Obviously )

I stained pancreatic cancer cells (Panc-1) for gamma-H2AX and got a large amount of foci in control cells (cells which have not undergone any genotoxic stress). I assumed Panc-1 cells are very genomically instable so their gamma-H2AX basal level is very high.

Did somebody get such a result?

Thank you!

What is the outcome of the treatment?

Hi,

If anyone can help me on finding out a list of different types of cancer cell lines that aberrantly overexpress the cell surface associated mucin1 (MUC1) proto-oncogene and the percentages of the MUC1 overexpression.

I have already known that MUC1 is aberrantly overexpressed in breast cancer (MCF7), non-small cell lung cancer (A-549) , prostate cancer (PC3), colorectal cancer (SW742) and pancreatic cancer (Aspc-1).

In addition, MUC1 overexpression has been demonstrated in hepatocellular carcinoma (Hep-G2), cervical cancer (Hala) and osteosarcoma (G292).

Thank you in advance!

It is my understanding that Tregs can influence development of autoimmunity through complex signaling pathways. Since they are often at high levels in progression of breast, ovarian and pancreatic cancers, I was wondering whether this is due to what's going on inside these tumors or their environment as a chemical process that may increase Treg production or differentiation, or whether the high levels of Tregs result from a patient's depleted immune system?

Can strategies be targeted selectively on reducing Tregs if they are interfering with a patient's response to therapy?

When a strain is not available on a repository such as Jax, how do I request an animal from another researcher? Is it as simple as sending them an email or is there another method to go about it that I am unaware of?

Are there any new discoveries on the biochemistry of pancreatic cancer that may serve as a marker? 

The particular application is for chemotherapy treatment of pancreatic cancer.  My understanding that the Bayesian approach is very popular with the FDA at present for certain applications.  I have a reasonable stats background, and some exposure to the Bayesian approach, but nothing related to clinical trial design.

Any suggestions most welcome.

Thanks,  Andy

using Panc1 and MiaPaCa2 cells

I have been trying to study the dose-response curve of paclitaxel (purchased from Santa Cruz) on pancreatic cancer cell lines (panc1, ASPC1, CFPAC-1)... I use drug concentrations starting from 0.01 nM to 10uM ... Though my drug range is wide enough, my cells weren't responding well, and my dose response curve remains more like a straight line.. Has anyone experienced like this? Could you guys please help me?

Pancreatic cancer is a worst type of cancer because it kills in a very short time, and there is currently no early detection method that I know where surgery can make a difference.

I'm working with this pancreatic cancer cell line and would be nice to know what the average cell diameter is. Can't seem to find it anywhere online. 

Specifically pertaining to colorectal and pancreatic cancer alongside other gastrointestinal malignancies, I was wondering what the experience has been in terms of data dictionaries, statistical support, size of databases and completeness of data. 

Hello Everyone,

I have a little doubt whether BxPC-3 pancreatic cancer cell line is KRAS Independent or Dependent?

I thinks its neither of it as it carries wild KRAS but need your confirmation.

Thanks

From what I understood is that the granulin protein has a role in the growth of the tumor to neighboring organs. I'd like to know more information about it to see if it's possible to inhibit it and prevent it from spreading.

I am trying to make a dissolve Gemcitabine prior to my experiments but when I look in the literature some people use water whereas other people use small concetration of DMSO which method is right and why?

Hello, 

I'm looking for an antibody against HNF4A that work for pancreas (that label HNF4A in the beta cells), I tried one from sigma and 3 from abcam, it works well in the liver but no signal in the pancreas.

If somebody can give me a antibody reference and/or some tips to make it work.

Thanks, 

Tom

Hi everyone. 

I would like to ask for your help :) Do you have some experiences with testing cell invasivity using Boyden chambers? I would like to coat my inserts with collagen I (rat tail) and Im having some troubles finding some universal protocol to start with (and have no senior in my group to ask for help). I found that finding the right collagen density is very important step and varry among different studied cell lines. Im working with crc and pancreatic cancer cell lines. Any suggestion how to coat the inserts and which concentration to use when work whith those? Thanks and have a nice day! 

Andrea

We hypothesis that chronic biliary pancreatitis is associated with pancreatic cancer initiation. But we are stucked in setting up an animal model.

Bile infusion method can be used in acute pancreatitis model, but not chronic pancreatitis model.

Caerulein injection is for chronic pancreatitis, but not biliary pancreatitis.

We have tried duct obstruction method by ligation the common duct of bile and pancreas. Due to high the mortality of the animals, we had the experiment paused.

Dose anybody have any experience about that?

Thank you very much for your help. We appreciate any comments.

I am working on a peptide which is designed to bind to a GLP1 receptor. Which cell line is most suitable to work on? I have tried MIN6 and RIN and have not got any positive results. I would like suggestions on which other cell line would be suitable. since GLP1 receptors are found in the gastrointestinal tract, which cell line related to the gastrointestinal tract would be suitable?

I am currently trying to generate stable cell lines using pSpCas9(BB)-2A-Puro (Addgene plasmid ID: 48139). The sgRNAs that were designed are successful in knocking down the protein of interest, however the knockdown achieved is only 10%. I am working with AR42J cells, which exhibits a poor viability and are relatively slow growing.

My question was whether I need to keep on selecting the cells under puromycin for several weeks or if the cells could be exposed to the drug for a shorter period of time (4-5 days) and that should be sufficient enough to kill untransfected cells/cells that have not undergone genome editing? Since the crispr/cas systems permanently edits the genome, I was thinking that the cells don't need to be cultured under the antibiotic selection.

Also is there any way I can increase the transfection efficiency so that a higher knockdown could be achieved with my transient transfections? I have tried lipofectamine 3000 which claims to provide a better transfection efficiency than others.

many hospital use endomicroscopic tools  ..laser, for deferent reasons for colon or pancreatic cancer

Hi,

I am treating pancreatic cancer cells with a range of growth factors. Surprisingly, we found a reduction in cell viability (measured by MTT), yet p-Akt was significantly up-regulated. 

The incubation time  was 72h

p-Akt is a pro-survival signal in cancer. However, I found a decrease in proliferation associated with it.

My questions: 

Is P-akt always associated with increased proliferation?

Would you suggest other cell viability assays or cell cycle analysis or looking at other pro-survival markers

Thanks

Do you agree that peripheral white cell counts (such as absolute lymphocytes count, ALC) was associated with cancer patients' outcome? Although lots of clinical studies confirm such relationships, but peripheral blood cells did not necessary correlate with immune cells in (peri-)tumor micro-environment. For peripheral ALC, we also did not have the ideals of actual subsets distribution of anti-tumor or immuno-suppressive lymphocytes....Regard to HCC, many "inflammation" index, such as ALC, NLR, PNI..was associated with patients outcome. So, I wonder such index only had roles of prognosticator, or it also provided "biological marker"  for future immuno-therapy.

There is a lot of debate in the efficacy of liver resection in metachoronus liver mets after whipple operation for pancreatic adenocarcinoma regarding better survival outcome than chemotherapy alone...what is your opinion?

Company name

Hi

I am working on a panel of pancreatic cancer cells studying the effects of cytokines on cell behaviour. BXPC3 especially grow well in RPMI media with 10% serum, however when I grow them on serum free, they die within 24h and during this time they form tubes. Because I am comparing the effect of the cytokine between different cell lines, I have to use consistent culture conditions. I found proliferation assay is done in serum-free in many publications.

So my question is it necessary to starve the cells? or as long as the cells are happy and I have control it would not be an issue?

Thanks

I want to design a study to assess whether an untested protein could be used as a biomarker for human pancreatic cancer.  I intend to do immunohistochemistry of this protein in pancreatic tumour tissue and normal tissue.  I am thinking of doing TMA and normal tissue microarray. Should I include any other assays like ELISA of body fluids and Western blotting for validating this protein?

Diabetes mellitus

but after three weeks no tumor was found in the liver either in large number of cancer cell , anyone know what could be the problem?

I am doing the research about checking the target molecules of FAM20C on the serectome of pancreatic cancer to induce the invasion and migration.

I want to determine the functions of ANXA2 in secretome on cell -induced FAM20C . So I would neutralizing ANXA2 on the pancretic cell which overexpressed FAM20C. 

But I cannot find detail the protocol for neutralization protein . Please help me if you know.

Thank you so much