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出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2016/01/24 11:42:51」(JST)

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Trypsinization is the process of cell dissociation using trypsin, a proteolytic enzyme which breaks down proteins, to dissociate adherent cells from the vessel in which they are being cultured. When added to a cell culture, trypsin breaks down the proteins which enable the cells to adhere to the vessel. Trypsinization is often used to passage cells to a new vessel.When trypsinization process is complete the cells will be in suspension and appear rounded.

For experimental purposes, cells are often cultivated in containers that take the form of plastic flasks or plates. In such flasks, cells are provided with growth medium comprising the essential nutrients required for proliferation, and the cells adhere to the container and each other as they grow.

This process of cell culture or tissue culture requires a method to dissociate the cells from the container and each other. Trypsin, an enzyme commonly found in the digestive tract, can be used to "digest" the proteins that facilitate adhesion to the container and between cells.

Once cells have detached from their container it is necessary to deactivate the trypsin, unless the trypsin is synthetic, as cell surface proteins will also be cleaved over time and this will affect cell functioning.^[1] Trypsin is inhibited by serum and the divalent cations calcium and magnesium, so serum is usually added to the container once cells have detached - this can be confirmed by observation under a microscope.

Trypsinization is often done to permit passage of the cells to a new container, observation for experimentation, or reduction of the degree of confluency in the flask by removal of a percentage of the cells.

References

^ Huang, H. L.; Hsing, H. W.; Lai, T. C.; Chen, Y. W.; Lee, T. R.; Chan, H. T.; Lyu, P. C.; Wu, C. L.; Lu, Y. C.; Lin, S. T.; Lin, C. W.; Lai, C. H.; Chang, H. T.; Chou, H. C.; Chan, H. L. (2010). "Trypsin-induced proteome alteration during cell subculture in mammalian cells". Journal of Biomedical Science 17 (1): 36. doi:10.1186/1423-0127-17-36. PMC 2873939. PMID 20459778.

External link

Cell Passage and Use of Trypsin (This link requires password access.)

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English Journal

Glycerol alters cytoskeleton and cell adhesion while inhibiting cell proliferation.

Dinsdale CJ, Mirza FM, Wiebe JP.SourceDept. of Zoology, University of Western Ontario, London, Canada.
Cell biology international reports.Cell Biol Int Rep.1992 Jul;16(7):591-602.
Our previous studies with various cell lines and human glioma cells showed that glycerol suppresses cell proliferation. In this study, we examined the effects of glycerol on the cytoskeleton, general morphology and attachment of Baby Hamster Kidney (BHK) cells. In glycerol-treated cells proliferatio
PMID 1516137

Preparing nuclei from cells in monolayer cultures suitable for counting and for following synchronized cells through the cell cycle.

Butler WB.AbstractA procedure is described for preparing nuclei from cells in monolayer culture so that they may be counted using an electronic particle counter. It takes only 10 to 15 min, and consists of swelling the cells in hypotonic buffer and then lysing them with the quaternary ammonium salt, ethylhexadecyldimethylammonium bromide. The cells are completely lysed, yielding a suspension of clean single nuclei which is stable, free of debris, and easily counted. The method was developed for a cell line of epithelial origin (MCF-7), which is often difficult to trypsinize to single cells. It works equally well at all cell densities up to and beyond confluence, and has been used with a variety of cells in culture, including 3T3 cells, bovine macrophages, rat mammary epithelial cells, mouse mammary tumor cell lines, and human fibroblasts. The size of the nuclei produced by this procedure is related to their DNA content, and the method is thus suitable for following cultures of synchronized cells through the cell cycle, and for performing differential counts of cells with substantial differences in DNA content.
Analytical biochemistry.Anal Biochem.1984 Aug 15;141(1):70-3.
A procedure is described for preparing nuclei from cells in monolayer culture so that they may be counted using an electronic particle counter. It takes only 10 to 15 min, and consists of swelling the cells in hypotonic buffer and then lysing them with the quaternary ammonium salt, ethylhexadecyldim
PMID 6496937