6 Steps for Successful in vitro Drug Treatment

6 Steps for Successful in vitro Drug Treatment

You might have seen one of the many anti-drug ads the 80s had to offer (including this delightful message from Robocop) and rightfully steered clear of drugs. But when it comes to biology, we use in vitro drug treatment for many experimental purposes, including testing anti-cancer treatments or synchronizing the cell cycle. If you are facing your first in vitro…

HIV-budding

A Beginner’s Guide to Lentiviral Transduction

The use of viral delivery systems to transduce cells for gene and protein investigations has become prominent over the last 20 years. In particular, the use of lentiviral vectors permits stable expression of your gene of interest. This is all possible with a little bit of nucleic acid magic. Lentiviruses (a genus of retrovirus) express reverse…

A brain working out to help get neurons firing

Tips for Getting Your Neurons Firing… Consistently

Primary cultures of rodent (rat and mice) neurons are widely used for disease modeling and studying cellular mechanisms in neurobiology, using a variety of techniques including neurobiology imaging. If you are in this field and need help with protocols and batch-to-batch variability of your dissociated primary rodent neurons, read further below. Also, consider watching several…

thp-1 cells

Mastering the Art of Growing THP-1 cells

Tissue culture can sometimes seem like a black art. Too careful—your cells go down. Not careful enough—your cells go down. A butterfly flutters its wings in the middle of the Atlantic Ocean—your cells go down. It’s annoying, it’s frustrating, and there are times (and I’m speaking from personal experience here) that you’ll end up chucking…

human clinical samples

Four Tips for Working with Human Clinical Samples

While using human clinical samples in your research can provide robust and heterogeneous results applicable to larger portions of the population, working with these samples presents its own set of challenges. Here are some tricks I have learned to help isolate and grow your cells of interest while eliminating stromal, blood, or other undesired contaminants….

anchorage-independent assays

How to Overcome Minor Issues in Anchorage-Independent Assays

Anchorage-independent assays test the ability of cells to grow independent of a solid surface. The assay is used to check the malignant potential of cancer cells. Cancer researchers generally do this experiment for any kind of confirmation of the oncogenic potential of an oncogene or a tumor suppressor in cancer cells. However, we do encounter…

sonication

Sonication – 7 Tips for Mastering the Art

Sonication is mostly used during preparation of protein extracts to help break apart the cell. Although most lysis buffers have buckets of detergent that lyse cell membranes, sonication just gives an extra hand in breaking everything apart. Sonication also breaks up, or shears, DNA in a sample—preventing if from interfering with further sample preparation. Have…

primary murine B cells

The Care and Keeping of Primary Murine B Cells

So you want to work with mouse B cells? Primary murine B cells are a difficult, yet fascinating system to work with and can help deepen your understanding of an immunological system. You can study many things with primary B cells, including: immune activation antibody production cell-cell interactions between immune cells and immune phenotype These…

Harvest Large Quantities of Secreted Protein with Hollow Fiber Bioreactors

Harvest Large Quantities of Secreted Protein with Hollow Fiber Bioreactors

Mammalian cell culture techniques are not simple, and culturing the cells requires a lot of maintenance as well as patience. In addition, doubling times compared to bacterial cells can take days instead of hours, which is most evident when contamination occurs. However, implementing small-scale hollow fiber bioreactors for culturing mammalian cells can save a lot…

dimensional cell culture

Cell Culture is No Longer Flat: Three Dimensional Cell Culture

Three dimensional cell culture mimics the extracellular matrix (ECM) that offers the structure and support for cells in vivo, thus creating the complex architecture and network required for cellular communication. For 3D cell culture beginners (or enthusiasts), the information available may seem overwhelming. It sure was for me. But it can be simplified. For example,…

How to See the Cell Cycle Through Your Microscope

How to See the Cell Cycle Through Your Microscope

Even in the most basic applications, fluorescence microscopy can be a very powerful technique. Simply put, the ability to actually see the biology you are interested in cannot be matched in directness. Often, the aim of fluorescence microscopy is to observe the effect of an experimental manipulation. Ultimately, you would like to know that the…

3 Carcinoma Cells, 2 Stromal Cells and a Partridge in a Pear Tree: An Introduction to Cell Co-Culture

The first thing you learn about culturing cells is proper aseptic technique and avoiding contamination. After that you’ll learn all the ins and outs of culturing your project’s specific cell line(s). What may not have been covered, is co-culturing, and I don’t just mean ethnic diversity in the lab! Co-culturing is the indirect or direct…

How to Culture Primary Human Bronchial Epithelial Cells

How to Culture Primary Human Bronchial Epithelial Cells

Human bronchial epithelial cells (HBECs) are a challenge to culture. As highly specialised cells that exist in carefully ordered multi-layered structures, they are especially fickle and finding optimum conditions to keep them happy is tricky. The cultures are also extremely sensitive to tiny changes in routine or environment. However, there are certain basic principles that…

Want to Increase Your Lentiviral Titers?  Focus on Your 293T Cells

Want to Increase Your Lentiviral Titers? Focus on Your 293T Cells

Producing lentiviral or retroviral vectors is theoretically fairly straightforward. However, anyone new to viral vector work is usually confronted with vast amounts of confusing information. It seems that anyone who has ever made a lentivirus has their own protocols and is adamant that their method is the best one to follow. In reality, there are…

express proteins

How to Express Proteins Across Kingdoms: Prokaryotes vs Eukaryotes

In the sci-fi novel Terminal World by Alistair Reynolds, a planet consists of zones with defined characteristics of matter interactions on a subatomic level. These conditions permit different levels of technology sophistication in various zones. For example, in the “Steamville zone” nothing more complicated than steam engines works – electronic schemes fuse irreversibly. Something like…

Top Tips on How to Prevent Cell Line Cross-Contamination

Top Tips on How to Prevent Cell Line Cross-Contamination

Recently we wrote an article about widespread cell culture contamination and how to detect it. This follow-up article will provide practical tips on avoiding cross-contamination in the first place. Be Cautious While Working The first way of cross-contaminating cultures is by accidentally mixing two cultures together, which may lead to an unintended co-culture or the displacement…