Cell line development and assurance of monoclonality are critical steps in the process of generating biopharmaceutical molecules, such as monoclonal antibodies. A cell line can be established following the isolation of a single viable cell robustly expressing the protein of interest. Cell Line Development. Cell line development plays a major role in developing robust, cost-effective biologic processes. Drug developers need rapid availability of high-performing mammalian cell lines for therapeutic protein production, and flexibility.
It is mandatory to procure user consent prior to running these cookies on your website. Cell Line Development — stable, high-yield, how to play bunker shots video clonal mammalian cell lines. Cell Line Development — stable, high-yield, what is cell line development clonal mammalian cell lines Rajesh Lohakare T Selexis offers comprehensive and rapid solutions for the development of stable, high-yield, and clonal mammalian cell lines that are used in recombinant protein drug manufacturing.
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Repositories of culture
Cell Line Development Workflow In order to generate high yields of recombinant protein products, stable cell lines such as CHO or HEK are typical vehicles of choice. In the case of monoclonal antibody production, the process of developing stable cell lines starts with transfecting host cells with recombinant plasmid DNA encoding the biotherapeutic molecule of interest. Oct 22, · Cell lines consist of transformed cell populations with the ability to divide indefinitely. This is typically due to immortalization in the laboratory or because the cell line is derived from a tumorigenic source from a patient or animal. Cell lines can be invaluable in research and have led to numerous important discoveries throughout medicine. Dec 30, · The cell line was developed by J.P. Jacobs in September after an abortion was committed on a physically healthy year-old woman for “psychiatric reasons.” In the Journal of Biological Standardization, Jacobs wrote, “The cells were derived from the lungs of a week-old normal male foetus at our laboratories in The family history was genetically normal; there was no Author: Carole Novielli.
Immortalized cell lines are critical for biomedical research, but establishing new lines can be tricky and frustrating. O n February 8, , George Gey of Johns Hopkins University isolated some cells from a cervical cancer biopsy and placed them into a petri dish with some medium. Unlike all of the other cells Gey and his colleagues had tested, these—from a patient named Henrietta Lacks— adapted to their new environment beautifully. Lacks died of her cancer eight months later, but her cells, dubbed HeLa, became the first immortalized cell line, capable of renewing itself in artificial culture indefinitely.
In the decades since their isolation, scientists have grown an estimated twenty tons of them. Meanwhile, researchers have identified numerous ways to transform primary tissues from humans and animals into immortalized cell lines, and now laboratory supply vendors and nonprofit repositories carry hundreds of lines specifically adapted for everything from protein production to virus propagation.
Embryonic and induced pluripotent stem cells may get more media attention, but ordinary somatic cell lines still form the backbone of biomedical research. The selection extends across a zoo of species. ATCC currently holds more than three thousand lines. The Coriell Institute for Medical Research in Camden, New Jersey maintains several thousand more, with an emphasis on human lines representing specific diseases.
Even with thousands of cell lines just a click or phone call away, though, scientists may still find the selection inadequate. The serpent fan explained that IBD triggers behavioral changes, followed by wasting, secondary infections, and death, and is a major problem in the pet snake trade. Veterinarians had no idea what caused it. Intrigued, DeRisi and Stenglein decided to see if a virus might be responsible. Working with snake owners and veterinarians, the team performed metagenomic sequencing and uncovered evidence of arenavirus infections in snakes with IBD.
The trouble started when Stenglein tried to grow the new viruses. Through their new veterinary contacts, Stenglein and DeRisi collected tissues from a boa constrictor named Juliet, which had died of lymphoma. To keep the problem manageable, he minimized as many variables as he could. For example, he used only one recipe for the cell culture media: minimal essential medium MEM —a liquid solution, originally invented by Harry Eagle, which meets the basic requirements for many cells.
These days a wide array of other media have been developed to help cater to specific needs, including sensitive and difficult-to-culture cells and lines from different species. Stenglein also incubated all of the plates at the same temperature, varying only the cell isolation method. The protocol that finally worked involved simply slicing tissues into pieces with scalpels, then immersing them in trypsin overnight.
The new lines have now propagated through multiple passages. Using these, the researchers developed a test for the new arenaviruses and used it on snakes with and without IBD.
The work showed a strong correlation between arenavirus infection and IBD, suggesting that the viruses may cause the disease. Stenglein now sees developing new cell lines as simply another laboratory technique that he could use in the future—though he has no immediate plans to do so. They did offer them to a repository, but were declined. Government funding for the repository ended 20 years ago, though, forcing ATCC to become more selective.
Now, the nonprofit organization only adds new lines for which they anticipate high demand and widespread scientific interest. The Coriell Institute still receives substantial federal funding, but focuses on human and nonhuman primate cells.
The institute maintains a trove of several thousand samples that range from umbilical cord blood to clinical isolates from patients with rare genetic diseases. Besides banking established cell lines, repositories are also at the forefront of creating new ones. The simplest way to create a new cell line is to modify an existing one, a common strategy when an established line already comes close to meeting the requirements.
Cells optimized to grow particular viruses or maximize recombinant protein production often come from such modifications. Establishing an entirely novel cell line can require more exotic techniques. While traditional cut-and-try methods such as those used by Stenglein may work, professional cell line developers constantly look for ways to accelerate the process.
For example, the large T antigen from SV40 virus, or the E6 and E7 oncogenes from human papilloma virus, can quickly turn a primary cell culture into an immortalized line. Originally developed in , the hTERT technique can yield cells that behave like primary cultures but propagate like immortalized lines.
Tumor samples often need little help becoming immortalized, having already acquired the ability to replicate indefinitely. For lymphoblasts, the easiest method is often infecting the cells with Epstein-Barr virus, which naturally transforms the cells but allows them to maintain much of their normal physiology. Tian advises researchers who think they need a new cell line to start by reviewing the literature and repository databases.
If no existing lines seem to come close, the next step is to decide on a general strategy for immortalizing primary cells. BioCat GmbH in Heidelberg, Germany also sells established cell lines and reagents for cell immortalization.
Alternatively, researchers can send their primary cells directly to BioCat and let the company develop the cell lines. Regardless of the specific techniques used to create a new line, the usual problems of any type of cell culture can arise, such as microbial contamination. Mycoplasmas are particularly hard to handle, as they can grow unnoticed in a culture and quietly ruin experiments.
Preventing contamination is even better, and incubator manufacturers have attempted to facilitate this through improved designs with features such as easier to clean surfaces, automatic decontamination cycles, and integrated air filters. Though clever techniques can sometimes speed the process, each cell line derivation problem will be unique, as Eric Donaldson discovered a few years ago. As a postdoctoral fellow with Matthew Frieman, assistant professor of microbiology and immunology at the University of Maryland School of Medicine in Baltimore, Maryland, Donaldson was looking for new coronaviruses.
Unfortunately, these viruses have been hard to isolate from their natural reservoirs. Donaldson thought a new bat-derived cell line might help. He quickly discovered that bat biologists are more reserved than the snake enthusiasts that Stenglein worked with.
He adds that he had to prove to bat biologists that he was interested in helping the animals rather than destroying them. Eventually, Donaldson and Frieman established a relationship with the Save Lucy Campaign , a bat conservation program based in Annandale, Virginia. Because bats can harbor serious human pathogens, including rabies viruses, the researchers worked with the tissues in a biosafety level BSL -2 plus facility with containment cabinets and restricted access.
Donaldson used a cell sorter to separate the cells of each tissue, and introduced hTERT and a mouse oncogene called BMI-1 into the cells to try to immortalize them. Most of the experiments, involving multiple tissues from 10 different bats representing four species, failed.
Creating new cell lines from three species is impressive, but bats are the most evolutionarily diverse order of mammals, and epidemiologists increasingly suspect that they may be major reservoirs for numerous emerging pathogens. Initially, Eckerle encountered the same barrier as Donaldson: bat tissue is hard to get. While working on that problem, she began practicing her tissue culture techniques on pig cells, establishing protocols to turn various primary tissues into immortalized cell lines.
She also developed a technique for rapidly freezing cells in the field. When she finally got some bat tissue, Eckerle and her colleagues created novel lines from the airway epithelia of two bat species representing the two major suborders, Yangochiroptera and Yinpterochiroptera. After carefully dissecting the tiny tracheae and establishing primary cultures, she used SV40 T antigen to immortalize them.
She and her colleagues are now establishing additional cell lines from other bat tissues and species. Inclusion of companies in this article does not indicate endorsement by either AAAS or Science, nor is it meant to imply that their products or services are superior to those of other companies.
Life science researchers who want to expand their capabilities beyond traditional cell counting instruments can now do so with the Countess II FL, a next generation, benchtop assay instrument with a modular design that broadens the number of applications on a single platform.
Designed with flexibility in mind, the Countess II FL can also be configured to use a full range of EVOS light cubes that provide more than 13 fluorescence color options. The instrument can operate with a reusable glass chamber slide to reduce the cost of consumables.
New Cell Culture Consumables offer a new dimension of safe, reproducible, and reliable cell culture work. Scientists and technical personnel in the field of cell culture have a strong need for easy, safe, and reliable products with improved handling that help prevent contamination. The latest products from Eppendorf deliver exceptional levels of product purity and security, as well as improved, ergonomic and safe handling of cell cultures and advanced protection against contamination.
Innovative technologies in Eppendorf Cell Culture Consumables deliver ultimate ease of use. Cell Culture Consumables Eppendorf. BrandTech Scientific announces the availability of inertGrade microplates for the cultivation of nonadherent cell lines, spheroids, and stem cells in a well plate format. These polystyrene plates are manufactured in one of the largest, most modern cleanroom facilities for laboratory disposable items in the world.
Rather than the more common treatments to enhance the natural hydrophobic characteristics of polystyrene, these new microplates are treated with a proprietary hydrogel. This creates a hydrophobic interface between the surface of the plastic and cellular material to inhibit cell and protein attachment. This unique low-binding microplate surface successfully suppresses the adhesion of a wide variety of adherent cell lines, can enhance the formation and maintenance of uniform spheroid cultures, and can inhibit early differentiation of stem cells or neurospheres.
InertGrade well microplates are available with round, flat, or curved bottoms in clear, white, black, and colored with transparent bottoms. The SNAP i. The power of IHC lies in its capacity to localize antigens within tissue samples, thereby identifying the cell types and subcellular compartments in which antigens are located.
Traditional immunohistochemistry methods may be subject to process variability; for example, the process typically requires a lot of manual slide handling, as well as the use of pap pens and pipettes. It decreases slide handling time and enables parallel processing of up to 24 slides at once, reducing slide-to-slide process variation without incurring the costs of automation. In addition, the system speeds wash steps and allows antibodies to be recovered and reused, saving researchers valuable time and resources.
The system produces robust and consistent staining, without causing tissue degradation or blotchy artifacts. Assisting researchers in gaining a deeper understanding of dynamic biological processes, the new cellSens imaging software version 1. Enabling effortless setup of complex acquisition sequences and protocols, the GEM presents an intuitive method to seamlessly control motorized hardware, delivering outstanding ease and efficiency for advanced live cell imaging applications. In cellSens 1.
Prioritizing the use of fast devices such as piezometric z -axis modules reaches a new level of imaging speed, with the freedom to choose the sequence of motorization movements. Enhancing the efficiency of time-lapse applications, the improved GEM also enables investigations into short-term dynamics and long-term sample evolution side by side. Visit Science New Products for more information. Newly offered instrumentation, apparatus, and laboratory materials of interest to researchers in all disciplines in academic, industrial, and governmental organizations are featured in this space.
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New products in Cell Culture Featured Product Countess II FL Life science researchers who want to expand their capabilities beyond traditional cell counting instruments can now do so with the Countess II FL, a next generation, benchtop assay instrument with a modular design that broadens the number of applications on a single platform. Search Jobs Enter keywords, locations or job types to start searching for your new science career.