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In this exclusive interview, we interviewed Dr. Carlemi Calitz, a postdoctoral researcher at Uppsala University, who worked in Dr. Femke Heindryckx’s group to understand how their team developed the most advanced 3D model to deal with hepatocellular carcinoma. We also learned how Calitz accidentally discovered the existence of metastases by observing "climbing" cells, and how Calitz hoped that her model could be used to help others study many forms of cancer, in order to improve treatment methods and stay in cancer research. Own mark.
Hepatocellular carcinoma (HCC) is a primary malignant tumor of the liver that often affects individuals with underlying chronic liver disease and cirrhosis. Liver cancer is the second largest cancer in the world and is usually diagnosed at an advanced stage when there are few treatment options. The Calitz and Heindryckx team is working to develop a realistic 3D model that scientists can use to help predict drug response and study this and other forms of cancer.
Calitz is facing how to better treat HCC positively by studying the sine curve. The sine curve and liver cells are the most basic structures of the liver. "The way the sine curve is structured is that we have a layer of endothelial cells. Under this layer, we will find stellate cells and hepatocytes," Calitz explained. "Once the liver is damaged, the stellate cells will migrate to the liver cells and begin to secrete type I collagen. This process will cause the liver to harden, leading to fibrosis, cirrhosis, and eventually HCC."
The team at Uppsala University used cell culture plates to help them visually observe the endothelial layer on top of the sample, where stellate cells and hepatocytes are embedded in a physiologically relevant hydrogel. Calitz explained: “The gel composed of stellate cells and hepatocytes is composed of collagen and fibrinogen, which causes changes in stiffness. Therefore, we can simulate fibrosis, liver cirrhosis, and complete HCC.” Calitz is based on these components. Developed her unique 3D model so that she can observe three different situations in one electroplating system. "If you only focus on one cell and ignore other cells, you can't fully understand what's really happening. That's why we need to target multiple cells at the same time to really understand the response to the drug."
3D cell culture technologies continue to receive widespread attention because they may provide more accurate tissue models. There is no doubt that 2D and 3D cell culture are related to their own benefits and frustrations. Many scientists question whether it is time to start the transition from 2D to complete 3D cell culture technology. Calitz explained that her model cannot be developed using 2D culture at all: "It may take seven months to prepare a mouse model for research. My model only takes three weeks and can last for a while. I am now in the laboratory. The model has been used for 25 days and is still fully feasible. This is cool.” Calitz told us that the ability to control the stiffness of the gel in 3D is also important: “By adjusting the stiffness, you can get more realistic and organ-like This will also have an impact on cell communication. It all comes down to cell communication, because this is how we solve the mystery of cancer, because cells interact by releasing certain molecules, proteins and enzymes.” Calitz also Point out that her 3D model can be cultured for about 25 days, while using 2D culture, a cell can only be cultured for 3 to 7 days. "Within three to seven days, there is no more space in the flask and you have to transfer the cells to a new flask. Therefore, the two-dimensional system is only suitable for acute studies. With 3D culture, this unwanted event will not happen. Cells can continue to grow and maintain viability." In 3D models, cells can maintain viability for a long time, making them ideal for acute and sub-chronic research before being transferred to animal models.
Despite the success of 3D cell culture, challenges still exist, such as long culture time, staining, imaging, and optimization. "In three-dimensional cell culture, there may be a lack of standardization. Some scientists perform cell culture for 7 days, while others require 11 to 18 days," Calitz continued. "The Celvivo team found that it takes 18 days for cells to recover from trypsinization. . This is important because the age of the sphere affects the results you get.” Calitz also elaborated on why her process still needs to be optimized: “Another problem we face is our process optimization, such as ratios and knowing how much we should add Content. This may require a lot of trial and error, which requires time, patience, and the need to collaborate with other experts."
The Calitz and Heindryckx team not only developed promising models to improve cancer treatment, but they also made surprising discoveries in the process. Calitz explained that her model is now called the metastatic model, which means that she can physically observe and study the metastatic factors associated with HCC. "I looked under the microscope and I saw the cells crawling out of the gel and falling into the medium." Like a real scientist, Calitz questioned why her cells came out of the gel, so strange Does the phenomenon prove to be a real transfer? She recalled: “The first thing I thought about was that this must be metastasis. I checked the literature and found some interesting papers about metastasis in other cancer models.” In order to find the answer, Calitz began to collect climbing cells. "I started to collect the cells in the gel. This is a difficult task because you can't simply remove the cells from the gel because this creates a sticky platform, which means I can't isolate RNA from the cells, so It takes time and planning." After the cells were isolated, Calitz performed multiple PCR tests and found that her cells expressed many epithelial-to-mesenchymal transcription factors and other known metastasis markers. This discovery can be used to explore metastatic cancer and has real research potential in the future.
Calitz and her team need the most advanced tools to help them achieve their research goals, and they have found a solution in Thermo Fisher Scientific technology. "Most of the equipment I buy comes from Thermo Fisher Scientific, and this includes all my PCR equipment, imaging and low-attachment plates, Gibco cell culture media and hepatocytes," Calitz said. "The equipment I bought from Thermo Fisher Scientific is really the best, especially the low-attachment plate. In just two days, we have been able to develop a beautiful and perfect round sphere in the middle of the plate. I will never I will use other plates." For Gibco cell culture media, Calitz said that she had never had a bad experience, and added: "Everything is of high quality, and I will not use any other media."
Looking to the future, Calitz revealed that her goal is to play a role in advancing cancer research. "I may not find a cure for cancer," she told us, "but if I can put a brick into the wall for others to build, this is what I want to do, if my 3D model is one of them, So this is what I hope to achieve." Calitz predicts that the future will be 3D: "I don't think we will completely eliminate 2D culture, but I believe 3D will soon become more important than ever."
Check out this guide to learn more about 3D cell model culture and analysis >>
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Thermo Scientific™ Nunclon™ Sphera processing...
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