Is the culture surface coated ?
No, it is not coated with any chemical substances, protein, gel, etc.
What is the difference between other three-dimensional cell culture methods ?
Two of the most popular 3D culture is, 1) by forming spheroids within matrix or gel, or 2) by forming spheroids suspended with Ultra-low attachment microplates. NCP is a ready-to-use microplate, which can form spheroids with a standard culture medium, and can be easily observed by standard microscope through the transparent bottom film, and has following advantages.
* Compatible with High-throughput
* Easy to observe through transparent bottom film
* High cell viability (compared to Ultra-low attachment microplates)
* Easy to collect cells
* Easy to change medium (spheroids attach to the plate, compatible with fluid medium)
What is three-dimensional (3D) cell culture? What is it advantage ?
Three-dimensional (3D) culture is a culture method to form three-dimensional multi-cellular cell cluster (spheroid). Cell morphology is different compared to conventional culture method using polystyrene cultureware (two-dimensional (2D) culture, monolayer culture). Multi-cellular spheroid is known to have closer characteristics to in vivo compared to monolayer cultured cells.
At its root, cancer is a disease of the DNA. But to cure it, we need to move beyond genetics and work together to uncover cancer’s deeper cellular chemistry, says Ronnie Andrews, President, Genetic and Medical Sciences at Life Technologies. Scientific American spoke with Andrews about this new approach to combat cancerin the 21st century.
How has our understanding of cancer changed in recent years?
We now consider cancer as a systems biology disease, not just a disease of genetics. Depending on what numbers you use, 12 to 18 percent of cancers are familial or hereditary—which means the rest of them are not. They are somatic, in that the mutations that cause the cancer arise during a person’s lifetime. So there’s something about people’s biology that either predisposes them to cancer or keeps them healthy. In the next four to five years we will be peeling back this onion, trying to figure out what it is about people that make them either more or less susceptible.
What does this mean in terms of finding a cure? Are we still really far away?
I’m going to give you my Wall Street explanation of how cancer develops. Let’s say the blueprint for a cancer is developed in a downtown Los Angeles architectural firm. But the cell doesn’t become cancerous unless the blueprint makes it to a manufacturing facility located in Newport Beach. So the architects give their blueprint to a courier who drives to Newport Beach and can get there one of many different ways—if a road is blocked off somewhere, he can take a different route. After the Human Genome Project, pharmaceutical companies realized that they may not be able to change the blueprint for cancer—as in, the genetic instructions for it—but they could potentially block the highway system so that the courier couldn’t get to the manufacturing center and deliver these instructions. By this I mean that it might be possible to keep the cancer genome from being transcribed—to prevent the “bad” proteins that create massive cell production from ever being made.
Can you walk me through how an individual cancer patient might get treated using this approach?
Let’s say you get a biopsy of a person’s tumor and identify a handful of cells of interest in that tumor. Then you do a whole genome sequence analysis of those cells and see all of the multiple potential mutational drivers of the cancer. At the same time, there are new proteomic tools coming out that will show us in real great detail and quantification not only what proteins are being produced, but where they’re being produced in the cell.
Once we have all this information, what do we do? Let’s go back to my analogy. What we want to do is make a Google map of the patient’s cancer cells. Then a doctor could figure out where the courier is at any point in time and pick drugs that stop the courier today by disrupting a particular biochemical pathway. And, after identifying future off-ramps that the courier might take, doctors could also prescribe drugs that would thwart the cancer in the future.
To find this perfect cocktail, you can imagine that a doctor would flip open an iPad, log on to a portal and pull down his patient’s information, which is password and HIPAA protected. Then he would relate the data from his patient to outcomes from similar patients from the past, whose details have been kept, anonymously, in a centralized database. He’ll be able to query the database and say, “show me 50 patients around the world that look like my patient at the genome and protein level, and now show me the top protocols that have allowed for the best survival rates for that patient.” This is the power of bioinformatics that we’re now all chasing.
This kind of approach requires scientists and doctors to share their data. Isn’t this unrealistic? I mean, scientists are notoriously protective of their data so they can publish it.
The reality is, in cancer, no single discovery is going to be a cure. So it’s time for us to become a little more selfless and to think about contributing to the greater good. Memorial Sloan-Kettering Cancer Center, for instance, may see 6,000 to 8,000 breast cancers next year, and MD Anderson Cancer Center might see 10,000. But there are hundreds of subtypes of breast cancer at the molecular level, so if these institutions don’t share their data, they will never collect enough of one cancer type to create a statistically powered data set allowing them to predict the best course of therapy for a given patient. At Life Technologies, we are creating a multi-company-sponsored program involving 20 institutions globally to do just this. We need to aggregate our data so that we can find solutions today instead of in 20 years—because in doing so, we will save thousands of lives.
What is the difference between “Cell line,” “Cell strain,” and “Cell type?”
When cells are isolated from a tissue to form a primary culture, assuming that the cells proliferate in vitro, a confluent monolayer or a dense cell suspension is formed. According to the traditional definition, the first harvesting and subculture of this cell population results in the formation of a cell line [Freshney, R.I. (1987). Culture of Animal Cells. A Manual of Basic Technique. (New York, Alan R. Liss, Inc.)]. This type of cell line has a finite lifespan, during which cells with the highest growth capacity will predominate, resulting in a degree of genotypic and phenotypic uniformity in the population.
Using this nomenclature system, a continuous cell line is a population of cells that has undergone a genetic transformation, resulting in indefinite growth potential. Continuous cell lines are usually aneuploid. In practice, continuous cell lines can be cultured through a very high number of subcultures, although some further genotypic, and therefore phenotypic, changes may occur at very high passage numbers. Immortalization can occur spontaneously, or may be virally- or chemically- induced. It should be remembered that the working definitions of these terms can vary between research groups. Many researchers do not use the term “cell line” to refer to any population unless it has undergone a genetic transformation.
A cell strain is a subpopulation of a cell line that has been positively selected from the culture, by cloning or some other method. A cell strain will often have undergone additional genetic changes since the initiation of the parent line. Individual cell strains may, for example, have become more or less tumorigenic than the established line, or they may be designated as a separate strain following transfection procedures.
The term cell type refers to all cells with a common phenotype, eg keratinocyte, melanocyte. Therefore keratinocytes isolated from a number of different donors are all the same cell type.
What does “normal” mean in the designation “normal human cell type?”
“Normal” means that the cells in question were isolated into primary culture from normal healthy tissue, rather than from diseased tissue. “Normal” also refers to a cell population that constitutes a cell line as opposed to a continuous cell line, according to the traditional definitions given above, since the cells have not been genetically altered and do not have indefinite growth potential.
Can a frozen ampule of cells be put back into liquid nitrogen and stored for any length of time?
In many cases, an ampule shipped in dry ice (-80°C) can be placed back into liquid nitrogen and the population recovered by rapid thawing at a later date. However, the viability may be reduced by such treatment, and for some sensitive cell lines, this may make recovery more difficult. The phenomenon is thought to be due to a change in the ice crystal structure within cells that occurs during the temperature shift. For this reason, we recommend that cells be thawed and placed into culture as soon after receipt as possible. It is best to minimize storage time at -80°C; that is, to use this temperature only for shipping. Celprogen does not warrant the viability of cells stored at -80C after shipments have been received.
How many passages can I obtain with Cell Applications’ primary cells?
Cell Applications uses the term “population doubling” instead of “passage” for describing the growth potential of the cells. A population doubling is a two-fold increase in the total number of cells in culture. A passage is the propagation of a cell population by subculturing from one vessel to 3 or 4 vessels. Because different researchers use different split ratios when they subculture, it is difficult to predict how many passages can be obtained with a particular primary cell. Most of Cell Applications primary cells are guaranteed for at least 15 population doublings (unless indicated otherwise in the product literature) when Cell Applications’ cell-specific growth media are used for the culturing procedures.