I'm going to study the anti-inflammatory effects of carvacrol and thymol in vitro using ELISA kits for TNF-a and IL-6. I have to use only monocytes? I'm going to use cells from patients with acute leukaemia and I will activate them with LPS.
Dear Ioanna,
You can use the cells indicated in the attached paper entitled " Human Vascular Endothelial Cells: A Model System for Studying Vascular Inflammation in Diabetes and Atherosclerosis". I have copied the paper abstract for quick view.
Curr Diab Rep. Author manuscript; available in PMC 2012 Jun 1.
Published in final edited form as:
Curr Diab Rep. 2011 Jun; 11(3): 193–202.
doi: 10.1007/s11892-011-0182-2
PMCID: PMC3311155
NIHMSID: NIHMS356778
Human Vascular Endothelial Cells: A Model System for Studying Vascular Inflammation in Diabetes and Atherosclerosis
Duygu Onat, David Brillon, Paolo C. Colombo, and Ann Marie Schmidt
Author information ► Copyright and License information ►
The publisher's final edited version of this article is available at Curr Diab Rep
See other articles in PMC that cite the published article.
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Abstract
The vascular endothelium is the inner lining of blood vessels serving as autocrine and paracrine organ that regulates vascular wall function. Endothelial dysfunction is recognized as initial step in the atherosclerotic process and is well advanced in diabetes, even before the manifestation of end-organ damage. Strategies capable of assessing changes in vascular endothelium at the preclinical stage hold potential to refine cardiovascular risk. In vitro cell culture is useful in understanding the interaction of endothelial cells with various mediators; however, it is often criticized due to the uncertain relevance of results to humans. Although circulating endothelial cells, endothelial microparticles, and progenitor cells opened the way for ex vivo studies, a recently described method for obtaining primary endothelial cells through endovascular biopsy allows direct characterization of endothelial phenotype in humans. In this article, we appraise the use of endothelial cell-based methodologies to study vascular inflammation in diabetes and atherosclerosis.
Keywords: Inflammation, Diabetes, Atherosclerosis, Adhesion, Apoptosis, Vascular biology, Endothelium, Endothelial function, Endothelial cells, Vein, Cell culture, Stem cells
Hoping this will be helpful,
Rafik
I have read papers that used HL-60 cells and THP-1 cells to study inflammatory markers. I have used both cells lines and they are both robust cells and can be stimulated to increase inflammatory cascade.
THP-1 cells worked well for me...
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If you are going to use blood samples from patients, why not use peripheral blood mononuclear cells, granulocytes, and monocyte-derived macrophages? This would give you maximal data from each sample.
Many thanks to all of you for your valuable replies. Carol i would prefer to take a cell line. All the papers i have studied used THP-1 cell line. But if i dont find it, what other cell line i could use?
Human umbilical vein endothelial cells, human coronary artery ECs, and human aortic ECs are commercially available. We have successfully used these primary cells to study inflammatory response and immunomodulation. Peripheral blood derived monocytes, neutrophils and T-lymphocytes can be used depending on your research objectives. If you are using immortalized cell lines it would be a good idea to validate the findings in primary cell cultures. Best wishes.
THP-1, HL-60, Monomac 6 are all good cell lines you could use, but it would be useful to try some PMBC experiments from 'normal' donors to compare with your patient samples.
All the suggested cells are suitable. But it will be better to choose a cell line depending what expectation you have from carvacrol and thymol. Do you want to see their effect on vascular inflammation?
Mono mac6 cells produce the most TNF alpha after LPS stmulation. I agree with Carol. THP1 cells are ok too.
If i cant use a cell line, could i isolate the peripheral blood mono-nuclear cells called buffy coat (PBMC) from blood using Ficoll? Im going to use blood from patients with leukemia and from healthy donors. Is it necessary to differentiate the cells with PMA?
Yes, but you will have to try keeping conditions of the experiment the same between repetitions: take the blood sample to the same donor avoiding circumstances which can alter the inflammatory responses (i.e. infections, allergies, drug prescriptions, etc) and using unstimulated blank for each chemical concentration tested. The above also applies if you going to use a certain "n" of patients/controls, except for the same donor part.
In regard to stimulation, you either use LPS or PMA as they both activate different monocyte intracellular pathway (reference pending).
Good luck
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