Reactive Oxygen Species (ROS) are reactive molecules and free radicals derived from molecular oxygen involved in cellular homeostasis. An excess of ROS production (e.g. exposure to environmental stress such as UV or heat exposure) causes significant damage to cell structures. In this post, let’s review the research tools for studying this process also known as Oxidative stress.
All-in-one system for iPS and hES gene editing
Today, I’d like to invite you to take a look at a highly efficient and useful kit, which brings together all the required components you need in a complete system for culturing and transfecting human pluripotent stem cells for gene editing.
The PluriQâ„¢ G9â„¢ Gene Editing System includes the G9â„¢ Maintenance Medium and G9â„¢ VTN Recombinant (vitronectin) plate coating for culturing human induced pluripotent (iPS) or embryonic stem (hES) cells in a manner that maximizes transfection by the included EditProâ„¢ Stem Transfection Reagent to transfect genome editing constructs. [Read more…]
SQSTM1 and autophagy
Immunofluorescence of monoclonal antibody to SQSTM1 on HeLa cell. [antibody concentration 10 ug/ml] (Cat No. H00008878-M01).
Autophagy was first described in the 60s, and it presents clear differences vs apoptosis. While apoptosis is a mechanism that kills the cells (apoptosis = self-killing), autophagy is more related to the orderly degradation and recycling of cellular components (autophagy = self-eating). [Read more…]
Proof that 2 = 5 ! Yes it does, in cell culture…
I have to admit that I never received the Fields Medal in Mathematics. Therefore, I won’t be able to develop this equation and prove that I’m right. However, what I can prove, is that in cell culture, 2 = 5. How is this possible?
Live cell imaging: the art of looking at biological processes in real time
Visualizing fixed cells and tissues only gives snapshots of cellular processes. To have a better insight into dynamic events ocurring in the cells or to vizualize interactions between various cellular components in real time (e.g., proteins, organelles, second messengers…), powerful microscopic approaches have been developed over the past decade. This post will review the recent live cell imaging probes developed by Goryo Chemicalsand available in Europe through tebu-bio.
Cost-efficient pre-optimized transfection reagents
The main challenge when choosing a transfection reagent is that we don’t know how it will work with our own cell type of interest. It is also time consuming to find the optimal conditions. Well, here’s the solution:  pre-optimised transfection reagents.
Pre-optimised transfection of pEGFP-N3 plasmid expressing eGFP under CMV promoter
They already cover more than 39 cell types including MEF, Caco-2, MCF-7, HepG2, Primary Macrophages, Huh-7 and many others.
Sharing our feedback on performances
Using a vector expressing the eGFP (pEGFP-N3) under CMV promotor, we assessed the transfection efficiency. Take a look at the example of results below. They will give you a pretty good idea of what you can expect.
Transfection efficiencies with pre-optimised Genjet reagents
Can we compare the pre-optimised Genjet?
Ok, now you can see we got good feedback. Still, will it be a better solution?
The answer is yes. The comparison with Lipofectamines, Fugene HD and Amaxa reveals that pre-optimised Genjet reagents allow high number of positive cells (dark green below).
You can see on right of each picture just above, that performances are even better with 10% serum, that would be much appreciated by the cells.
Furthermore, this quality is associated to low price. Check it for yourself!
Two top ways to success with knock-out
In March 2016, Mark J Osborn et al published in Molecular Therapy a major article for genome editing (doi:10.1038/mt.2015.197), about knock-out of CD3 in human T-cells. The goal is to improve T-cell-based immunotherapies to fight tumours using engineered allogenic T-cells from healthy donors. It is a very good example of how CRISPR-CAS9 can help medecine. And even if you are not very comfortable with CAR T-cells and the treatments of malignancies, I would recommend you read it and especially take a look at figure 2. Indeed, dear friends of genome editing, the authors made a clear and fair comparison of several KO strategies, covering all the main options. Thus, it is not only a major step for anti-tumour treatments but it is also an excellent overview that reveals the best approaches. So, before reading this post any further, you might like to read the article mentioned above. [Read more…]
Still using MTT or WST-1 for cell counting?
If the answer is yes, I am sure you’ll be interested to learn more about WST-8 and our Cell Counting Kit-8 (CCK-8). If the answer is no, and you’ve already switched to WST-8, just have a look at our price list… you might be surprised!
How to choose the best 3D technology for your assay?
Cell culture models using 2D substrates have provided important conceptual advances in understanding the biology of cells. However, cells grown on flat 2D surfaces can differ substantially from physiological environments. Animal models provide a useful tool to study biology in a physiologically relevant environment. However, animals models are expensive, time-consuming, use a significant amount of test material, and do not always provide a useful extrapolation to humans. In vitro 3D cell culture models bridge the gap between the two, allowing the study of human cells in a physiologically-relevant environment with the convenience and speed of an in vitro model. [Read more…]
From RNA-derived iPS Cells to Retinal Cells
Human induced pluripotent stem (iPS) cells and cells differentiated from iPS cells have widely been used for in vivo models human disease progression. Jason Meyer, of Indiana University Purdue University Indianapolis, uses iPS cell-derived models to study retinogenesis and retinal disease. Two recent papers from his lab highlight the benefits of using Stemgent’s RNA reprogramming technology to enable robust differentiation of iPS cells to the retinal lineage (1, 2). RNA reprogramming technology was chosen in order for these studies to ensure that no vestiges of the reprogramming vectors were retained by the cells or integrated into the genome.