Measuring cell count and viability in microcarrier cultures
– with NucleoCounter® automated cell counters
Microcarriers – a scalable culture system for adherent cells
Figure 1. Microcarrier bioreactor. Microcarriers offer a convenient method for growing adherent cells in bioreactors.
Scaling up cell and virus production can be challenging. Cell culture flasks are not feasible for industrial scale production when a thousandfold increase in production volume is needed. Microcarriers offer a convenient method for growing adherent cells in bioreactors. They serve as a scaffold that adherent cells attach to, allowing them to proliferate while a bioreactor keeps the cell-microcarrier complex freely suspended in the media. Therefore, adherent cell lines are grown like suspension cells, simplifying scaling and allowing existing resources to be leveraged for process optimization and production.
Technical Note (.PDF):The NucleoCounter® method for measuring cell count and viability in microcarrier cultures
Figure 2. NucleoCounter® instruments. The NucleoCounter® method for measuring cell count and viability in microcarrier cultures.
A NucleoCounter® detects cells by staining cell nuclei. The cells are made visible using the fluorescent dye DAPI, which is highly specific to DNA, giving accurate detection of cell nuclei even in the presence of cellular debris. Cell sampling, fluorescent staining and counting chamber loading are combined into a single workflow by the unique Via1-Cassette™, which is loaded into a NucleoCounter® NC-200™ or NC-3000™ , together calculating total cell count and viability.
Save time when counting cells growing on microcarriers
A comparison of the traditional trypsin digestion method and the NucleoCounter® workflow shows that the NucleoCounter® removes several centrifugation, pipetting and incubation steps. The entire cell counting process can be completed in less than five minutes.
Traditional trypsin digestion
and manual counting
The NucleoCounter® method
Literature
- Lam AT, Chen AK, Li J, et al., (2014), Conjoint propagation and differentiation of human embryonic stem cells to cardiomyocytes in a definedmicrocarrier spinner culture., Stem Cell Res Ther, Sep 15;5(5):11010.1186/scrt498
- Lam AT, Li J, Chen AK, et al., (2014), Cationic surface charge combined with either vitronectin or laminin dictates the evolution of human embryonicstem cells/microcarrier aggregates and cell growth in agitated cultures., Cell Therapy, Jul 15;23(14):1688-703, 10.1089/scd.2013.0645
- Heathman TR, Stolzing A, Fabian C, et al., (2016), Scalability and process transfer of mesenchymal stromal cell production from monolayer to microcarrierculture using human platelet lysate, Cancer Research, Apr;18(4):523-3510.1016/j.jcyt.2016.01.007
- Heathman TR, Glyn VA, Picken A, et al., (2015), Expansion, harvest and cryopreservation of human mesenchymal stem cells in a serum-free microcarrierprocess., Biotechnol Bioeng, Aug;112(8):1696-70710.1002/bit.25582
- Lam AT, Li J, Chen AK, et al., (2015), Improved Human Pluripotent Stem Cell Attachment and Spreading on Xeno-Free Laminin-521-CoatedMicrocarriers Results in Efficient Growth in Agitated Cultures, Biores Open Access, Apr 1;4(1):242-5710.1089/biores.2015.0010
- Chen AK, Chen X, Choo AB, et al., (2010), Expansion of human embryonic stem cells on cellulose microcarriers., Curr Protoc Stem Cell Biol., Sep;Chapter 1:Unit 1C.1110.1002/9780470151808.sc01c11s14
- Marinho PA, Vareschini DT, Gomes IC, et al., (2013), Xeno-free production of human embryonic stem cells in stirred microcarrier systems using a novelanimal/human-component-free medium., Tissue Eng Part C Methods., Feb;19(2):146-55.10.1089/ten.TEC.2012.0141
- Lecina M, Ting S, Choo A, et al., (2010), Scalable platform for human embryonic stem cell differentiation to cardiomyocytes in suspended microcarriercultures., Stem Cell Res Ther, Dec;16(6):1609-19.10.1089/ten.TEC.2010.0104
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