Genetic Engineering & Biotechnology News

JUL 2017

Genetic Engineering & Biotechnology News (GEN) is the world's most widely read biotech publication. It provides the R&D community with critical information on the tools, technologies, and trends that drive the biotech industry.

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Genetic Engineering & Biotechnology News | | JULY 2017 | 9 anticancer drug screening. 18 Consequently, primary cells are increasingly becoming the fo- cal point of 3D cell culture in cancer biology. 19 In cancer research, many studies on 3D cultures have used established cancer cell lines. However, more recently, using patient-derived primary tumor cells instead of cell lines has generated advanced, more biologically repre- sentative 3D models of cancer to aid drug dis- covery and research. 3 For example, one study based on a 3D model of primary human adult lung cancer-associated fibroblasts (LuCAFs) and human bronchial epithelial cells (HBECs) found that LuCAFs alter HBECs by modify- ing biochemical signals conveyed through the extracellular matrix. 20 A preliminary study by Lonza has also demonstrated that human mammary fibroblasts and human mammary epithelial cells could be successfully co-cul- tured (using the RAFT™ System) to produce a reliable multicellular model of breast cancer that could be used to conduct anticancer drug screening studies. Although using primary cells in 3D cell culture technology has not yet been opti- mized, it is recognized that its major ad- vantage is the ability to use the same tumor model in vitro and in vivo for cancer research and drug screening. For example, Kondo et al. developed a new method for culturing primary colorectal cancer cells. Using this method, the investigators maintained cell-cell contact throughout the culture process, and they showed that these cultured cells could be used for the evaluation of chemosensitiv- ity and signal pathway activation in cancer cells from individual patients. 21 A 3D-tumor model system using primary patient-derived cells could, therefore, promulgate discoveries in cancer research and in early-stage drug dis- covery for personalized drug programs. Primary Cell Culture: Tips and Tricks There are clearly many compelling reasons why primary cells should be used instead of cell lines for research, but their adoption has been gradual because of the widespread belief that they can be difficult to culture. Yet, prima- ry cells are just as easy to culture as cell lines, and simply require adherence to the specific protocols and commercially available growth media kits that are often provided by suppli- ers. Here are Lonza's top tips and tricks to ensure the successful culture of primary cells. 1. Before cells are cultured, they should be prepared correctly. The number of cells that will be needed (and thus the number of flasks) should be calculated beforehand, and the cells should be kept in liquid nitrogen for as long as possible before they are thawed 2. Cells should be thawed quickly (i.e., in no more than 2 minutes) because they can be harmed if thawing takes longer. Similarly, once the cells are in the medium, they can be temperature sensitive, so repetitive warming and cooling should be avoided. 3. After culture seeding and cell growth are established, cell proliferation should be stopped once the cells reach 70–80% conflu- ency. Be careful not to reach 100% confluen- cy, as this will make the cells enter senescence. 4. To dissociate the cells from the culture, cells should be washed with trypsin at room temperature. Because this process can be harsh, the cells should be monitored carefully through a microscope (Versene-assisted de- tachment is a milder alternative to trypsiniza- tion, if required). Afterwards, a trypsin neu- tralizing solution (again at room temperature) should be used to fully inactivate the trypsin. 5. If contamination is suspected for any rea- son, the culture should be checked regularly. Conclusion New three-dimensional cell culture sys- tems are driving forward research in several key biomedical fields, including cancer biol- ogy and drug discovery. The cell types avail- able for use in cell culture are either cell lines or primary cells. Increasingly, however, bio- logically relevant primary cells are becoming the preferred choice because of the various advantages they have over cell lines. Specifically, although cell lines are the familiar choice, they require authentication prior to use, are likely to produce unreliable and inconsistent results, and could potential- ly cause increased costs from failed animal tests and clinical trials in drug development. In contrast, primary cells are showing great promise for biomedical research. For exam- ple, they are producing biologically represen- tative 3D models of cancer to aid anticancer drug discovery. As such, using primary cells together with 3D cell culture systems could lead the way in advancing research and drug discovery, potentially helping to combat can- cer and other life-threatening diseases. Simple • Convenient • Efficient • Safe Lenti – Experience the Beauty! Focus on discovery, not DNA delivery Lenti Particles and Plasmids are Both Available Lenti-shRNA Lenti-ORF Lenti Packaging Kit © 2017 OriGene Technologies, Inc. All Rights Reserved. Drug Discovery Tutorial Lubna Hussain is senior global product manager at Lonza. Website: References available online.

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