According to the GE Healthcare publication "Microcarrier Cell Culture," a sample of culture of cells grown on microcarriers is placed in a test tube and allowed to settle. The medium is poured off and replaced with a trypsin and calcium solution. The tube is incubated while agitating, the microcarriers are allowed to settle and the medium is transferred to a second tube. The microcarriers in the first tube are washed and then pooled with the supernatant from the second tube. The suspension is spun, or centrifuged, and the supernatant is discarded. Trypan blue stain is added to mark to dead cell and cellular fragements, and viable cell counts can be made.
The addition of microcarriers to cell cultures is valuable for the study of endothelial cell biology, according to a study first describing this process in the February 1984 issue of "Laboratory Investigation." The researchers suggested a method for the selection of endothelial cells from mixed cell cultures using microcarriers in a migration transfer technique. Briefly, microcarriers were added to the cell cultures as a framework, on which the cells can multiply. In order to separate the cells, trypan blue stain was added. The stain did not attach to the microcarrier beads and made the selection of cells possible. In addition, the authors discovered that trypan blue stain attached only to dead cells and cellular fragments. Hence, the cells that did not stain were viable.
An article appearing in the December 1995 issue of "Histochemistry and Cell Biology" described a novel system to identify the cells involved in angiogenesis, which is the creation of new blood vessels as seen in tumor formation. The researchers developed a microcarrier-based cultivation system that uses trypan blue staining. This process allowed the researchers to observe the growth characteristics of several cell types. Trypan blue staining only enhanced the observation and did not have any effect on the microcarriers.
A study in the March 2001 issue of "Cytotechnology" detailed the characterization of cell growth and death in two types of media using microcarriers and trypan blue staining. The results of this study indicated that trypan blue staining was achieved using both media. The study concluded that trypan blue staining and the use of microcarriers could not determine whether cellular death was due to apoptosis, or genetically-programmed cell death, or necrosis, which describes the death of tissue by other means.