NucleoCounting provides a reliable and accurate approach to determine the cell count and viability in microcarrier cultures. Two reagents are used in this method, and these reagents quickly release the nuclei from the microcarriers.
The nuclei released are placed in a disposable Via1-Cassette™, which, in turn, is placed inside the NucleoCounter™. This instrument automatically carries out fluorescent staining, image acquisition, and image analysis. It takes less than 5 minutes to complete the entire process and can be carried out without using pipettes.
Microcarriers: A scalable culture system for adherent cells
Figure 1. Microcarrier bioreactor. Microcarriers offer a convenient method for growing adherent cells in bioreactors.
It can be difficult to scale up the production of cells and viruses. When there is a need for high production volumes, cells culture flasks are not a viable option for industrial scale production. Microcarriers make it easy to grow adherent cells in bioreactors, and they act as a scaffold to which adherent cells can bind to.
This enables the adherent cells to proliferate easily, while the cell-microcarrier complex is allowed to suspend freely in the media by a bioreactor. Therefore, adherent cell lines are cultured like suspension cells, simplifying the scaling up process and enabling the current resources to be used for production and process optimization.
The NucleoCounting method fluorescently stains cell nuclei in order to detect the cells. The fluorescent dye DAPI, which is very specific to DNA, is used to label the cells and it allows the cell nuclei to be accurately detected even when cellular debris is present.
The unique Via1-Cassette™, which is inserted into a NucleoCounter NC-200™ or NC-3000™ instrument that measures the total cell count and viability, combines fluorescent staining, cell sampling, and counting chamber loading into a single workflow.
NucleoCounting saves time when counting cells in microcarrier cultures
During optimization and monitoring of large-scale bioproduction, cell count and viability are very important parameters. Traditionally, cells that are cultured on microcarriers are counted using a multistep method, where trypan blue staining and trypsin digestion are used. However, this method is not only inaccurate, but also takes a long time.
The NucleoCounter workflow demonstrates that the NucleoCounter™ eliminates several pipetting, incubation, and centrifugation steps when compared to the conventional trypsin digestion method. In less than 5 minutes, the entire cell counting process can be completed using the NucleoCounter™ instrument.
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About ChemoMetec A/S
Founded in 1997, Chemometec specialises in the design, development and production of high quality instruments using patented technology for a wide range of applications in cell counting and evaluation.
Chemometec work closely with life science companies, research institutes, universities, hospitals, specialist clinics and a wide range of food and beverage manufacturers, matching our technical development expertise to customer needs - with quality results, reliability, cost-efficiency and ease-of-operation as our guiding principles.
Chemometec support a growing customer base worldwide with responsive technical services and the ready availability of a wide range of consumables.
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