Welcome to our complete package of video tutorials on cell culture technique. All commonly used procedures in the cell culture lab are described and discussed here.
We not only follow the rules for Good Cell Culture Practice, we also show, in great detail, how good practices should actually be implemented. Topics of the tutorials start from how to get ready for the cell culture lab, then we talk about cells, and everything connected to cells, such as cell culture medium, cell morphology, cell confluency, cell types, and cell line misidentification.
There is also an entire section dedicated to different kinds of vessels such as tubes, but also for the cultivation of cells, such as flasks and plates. We discuss passaging, counting and freezing cells, and we talk about contaminations in the cell culture lab. Of course, we also talk about lab equipment, in particular about pipetting with various pipettes. Ultimately, vessels, equipment and the way we treat our cells have a direct effect on them and on the data we generate.
The tutorials include common mistakes, common misconceptions, food for thought, but also good habits, opinions, tips and hacks, and do’s and don’ts.
Who will benefit
The tutorials can be watched in class, after class or before starting a practical course, or before starting a bachelor, a master or a PhD programme. They can be watched before starting to work in a cell culture lab, or before switching to a different workplace that happens to be a cell culture lab.
We hope the videos encourage you to discuss why things are done a certain way in the cell culture lab, and where things can still be improved. Regarding the order of the videos, we recommend a certain order, but, of course, you are free to choose a different order if that makes more sense to you and your situation.
In this video, we talk about recommendations for biosafety level 2 labs, and how you should prepare yourself for it.
Check out our Learning Toolbox Lab Support 1 (PDF for download) on how to get ready for the cell culture lab.
The empty space in the sterile workbench can be the reason why your experiment goes well or why it goes terribly wrong. Experienced cell culture people divide the space in the hood into functional areas, and all of a sudden, a tube, a rack, a medium bottle and a pipette become part of a superordinate structure. If this happens, many mistakes are less likely to happen, and the work-flow is one of a different kind.
When you start working in the sterile work bench (but even later, when you are more advanced), keeping in mind what is actually sterile, and what may be clean, but is not really sterile, can be a challenge. This video will help you to remember this literally small, but hugely important difference.
We do not only want you to become a well-rounded, confident scientist or lab technician, who is known for sound and solid data and scientific integrity. No, we also want you to be the person that has a pleasant aura, who is never rude or reckless, who is known to help others in any situation in the lab, and who is known to be trustworthy and to speak up whenever needed. We want others to think that you are the most likeable scientist or lab technician they ever met.
There are distinct differences between finite and continuous cell lines. If you work with cells, you should know these differences, and why they play a role.
In this video, we will discuss the differences and consequences of using finite or continuous cell lines in greater detail.
In this video, we will talk about different ways of isolating cells from a tissue and getting primary cells first and then establishing a finite cell line later in the process.
Of course, we cannot go too much into detail here, such as specific enzymes or enzymatic mixtures, coatings or specific methods to characterize the cells you isolated. But if, for example, one general method does not work, after trying everything else, you can try an entirely different approach to get your cells out of your tissue.
In this video, we will walk through a typical scenario of expanding a finite cell line. Cells of a finite cell line can only undergo a limited number of population doublings, which means, at a given split ratio, you can only passage your cells a limited number of times. So, you need to know the growth characteristics of your cells and plan everything strategically. Otherwise, you run the risk of not having enough cells for your experiments before your cells reach replicative senescence.
In this video, we will walk through the process of expansion of a continuous cell line. We will start at the time point, when you receive an original cell vial from an official cell collection. What are the steps that you have to take, so that you can maximize the cell yields from that single cell vial?
At the same time, we will ensure maximum reproducibility and standardization of our experiments.
We will also briefly discuss common mistakes in this process. Please keep in mind that the number of vials of the original cell bank may vary depending on the cell type. For some cell types you may consider a smaller number of vials for the original cell bank.
In the cell culture lab, we tend to forget that cells are not within their natural surroundings. Actually, many of the conditions, cells are cultivated in, are totally alien to them. Let’s discuss the significance of these not so fitting cell culture conditions, and how we - and our cells in culture - can deal with them.
In our experience, telling the confluency of a culture can be difficult for beginners. But this is nothing to be worried about. In this video, we will show you different ways of estimating cell confluency. After watching this, give yourself a little time, and with consistent practise you will manage to estimate cell confluency without any problems. Also check out our Learning Toolbox Lab Support 3 (PDF for download) with many examples of cell confluency.
It seems that cell culture people talk about cell confluency all the time. “Ugh, these cells are too confluent!”, or “Oh, no! What’s wrong with these cells?! They are still not confluent!” But what does it mean for the cells to be in a confluent, or not-so-confluent culture? Are they happier in one or the other culture? Let’s see if and why cell confluency matters! Also check out our Learning Toolbox Lab Support 3 (PDF for download) with examples of cell confluency for a jump start.
Passaging or splitting cells seems to be what cell culture people do all day long. But when do you actually passage your cells, and why? How many of the cells do you reseed? And what are you supposed to do with the remaining cells? We hope to answer all these questions in this video.
Enzymatic detachment is the most commonly used method to detach adherent cells from flasks and plates. But if things go wrong, these enzymes and some other factors and conditions during passaging can cause harm to the cells. So, let’s discuss a few factors that are important to consider during enzymatic detachment of your cells. Your cells should make it through the passaging procedure smoothly and without much damage.
In this video, we will go through a typical procedure of passaging a culture step-by-step with enzymatic detachment. We will start from the moment, when you take your cell out of the incubator, until we have our cell suspension of detached cells. Check out our Learning Toolbox Protocol 3 (PDF for download) on passaging adherent cells.
There are many things that can go wrong during passaging your cells. At the same time, many factors can be optimized in the process of cell passaging in order to lower the stress level for your cells - and for you! Let’s explore some of these factors!
In this video, we briefly go through the two basic methods of passaging suspension cultures.
We want you to become really comfortable with the term “split ratio”. Too much can go wrong, if split ratios are not given the attention they deserve. Cultures of cells can rise and fall, depending on their split ratio! Check out our Learning Toolbox Calculations 1 (PDF for download) on calculating cell numbers for passaging cells.
Here we will look at general differences between counting cells with an automated cell counter and manual counting with a hemocytometer. Why bother with manual counting if it is so much easier using an automated cell counter?
Manual cell counting can be a challenge. Clumpy cells, too many cells, too few cells, trypan blue killing your cells, you name it. All these issues should be addressed while counting, and managing them well will make you more and more comfortable counting your cells manually.
Let’s walk through the process of counting cells manually with a hemocytometer, together. We will go through the process step-by-step, pointing out typical pitfalls and common mistakes. We will show you, what you should expect to see under the microscope, and how the cells should look that you will count, and which ones you shouldn’t count. After watching this video, you will know, how to use the hemocytometer, and you will be able to distinguish between live and dead cells.
Here we have an example, of how you calculate your cell concentration after counting them. Not very exciting, we know, but it may save you some time in the beginning.
Check out our Learning Toolboxes Calculations 1 and 2 (PDFs for download).
In this video, you will get a collection of useful tips and hacks for manual counting. For example, what to do, when your cell suspension is not drawn under the cover slip of the hemocytometer, or how you can speed up your counting significantly. We will also talk about some safety issues, for example for handling trypan blue.
In this video, we talk about the theory behind the cryopreservation of cells. We will look at what happens to the cells during freezing and you will get a brief overview of the entire process. We will also talk about the cooling rate during freezing, and discuss why this is important.
In this video, we talk about the advantages and disadvantages of storing cells immersed in liquid nitrogen versus storing them in the vapour phase.