Wikipedia preview

出典(authority):フリー百科事典『ウィキペディア（Wikipedia）』「2013/12/22 18:12:16」(JST)

wiki en

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[edit]

^ 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. edit

External links[edit]

A suggested protocol for trypsinizing cells
Cell Passage and Use of Trypsin

UpToDate Contents

全文を閲覧するには購読必要です。 To read the full text you will need to subscribe.

1. 膵外分泌機能検査 pancreatic exocrine function tests
2. 小児および思春期における慢性膵炎の原因 causes of chronic pancreatitis in children and adolescents
3. 食物アレルゲンの分子機能 molecular features of food allergens
4. 急性膵炎の発症機序 pathogenesis of acute pancreatitis
5. 急性膵炎の臨床症状および診断 clinical manifestations and diagnosis of acute pancreatitis

English Journal

Human Keratinocytes' Response to Injury Upregulates CCL20 and Other Genes Linking Innate and Adaptive Immunity.

Kennedy-Crispin M, Billick E, Mitsui H, Gulati N, Fujita H, Gilleaudeau P, Sullivan-Whalen M, Johnson-Huang LM, Suarez-Farinas M, Krueger JG.SourceLaboratory for Investigative Dermatology, Rockefeller University, New York, New York, USA.
The Journal of investigative dermatology.J Invest Dermatol.2011 Sep 1. doi: 10.1038/jid.2011.262. [Epub ahead of print]
In the early stages of wound healing, keratinocytes (KCs) become "activated" and release inflammatory molecules such as IL-1 and IL-8, which are linked to innate immune responses and neutrophil recruitment. It is unclear, however, whether KCs release molecules linked to adaptive immune responses, e.
PMID 21881590

Biotinylated quercetin as an intrinsic photoaffinity proteomics probe for the identification of quercetin target proteins.

Wang RE, Hunt CR, Chen J, Taylor JS.SourceDepartment of Chemistry, Washington University, St. Louis, MO 63130, USA.
Bioorganic & medicinal chemistry.Bioorg Med Chem.2011 Aug 15;19(16):4710-20. Epub 2011 Jul 13.
Quercetin is a flavonoid natural product, that is, found in many foods and has been found to have a wide range of medicinal effects. Though a number of quercetin binding proteins have been identified, there has been no systematic approach to identifying all potential targets of quercetin. We describ
PMID 21798748

Japanese Journal

Proteome analysis of silkworm : 2. Hemolymph

KAJIWARA Hideyuki,IMAMAKI Atsue,NAKAMURA Masatoshi,MITA Kazuei,XIA Qingyu,ISHIZAKA Masami
Journal of electrophoresis 53(2), 27-31, 2009-06-01
… Prominent spots on 2D-gel were analyzed after trypsinization and proteins were identified by Mascot and Sequest using the amino acid sequence data obtained from the genome sequence of silkworm. …
NAID 10025192018

Streptolysin-O Treatment of Fetal Fibroblasts Improves Cell Fusion and In Vitro Development of Porcine Nuclear Transfer Embryos

NARUSE Kenji,QUAN Yan-Shi,KIM Baek-Chul,CHOI Su-Min,PARK Chang-Sik,JIN Dong-Il
The Journal of reproduction and development 55(3), 236-239, 2009-06-01
… In experiment 2, fetal fibroblasts were treated with 200 ng/ml SLO for 50 min at the confluent or single-cell stages of the nuclear transfer protocol (i.e., before and after trypsinization, respectively), and the in vitro development of the resulting porcine reconstructed embryos was investigated over 6 days in culture. … Oocytes receiving cells treated with SLO post-trypsinization showed higher fusion and blastocyst formation rates compared with those receiving untreated cells. …
NAID 10024942132