Abstract | AIMS AND BACKGROUND: As a powerful technique allowing analysis of large numbers of cells, fluorescence-activated cell sorting (FACS) is used more and more widely. For FACS analysis, adherent cells are usually detached by trypsinization, followed by centrifugation and resuspension. However, trypsinization can cut off some receptors from the cell surface like fine scissors, which will affect the accuracy of FACS results. Though non-enzymatic methods such as citric saline buffer have been used to determine cell surface receptors, how much of the receptors is cut off by trypsinization has been rarely studied. This work aimed to investigate whether different methods of detaching adherent cells could affect the detection of cell surface receptors. METHODS: RESULTS: Different methods of detaching adherent cells could significantly affect the detection of TRAIL receptors. Compared to the conventional trypsin- EDTA group, the non-enzymatic group showed a 3.42-fold increase in the mean fluorescence intensity index of DcR HepG2 and a 1.25-fold increase in DR Huh 7 (P <0.05). However, the viability, cell cycle and apoptosis of these cells were not affected. CONCLUSIONS:
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Authors | Bo Zhang, Hong Shan, Dan Li, Zheng-ran Li, Kang-shun Zhu, Zai-bo Jiang, Ming-sheng Huang |
Journal | Tumori
(Tumori)
Vol. 98
Issue 6
Pg. 800-3
(Nov 2012)
ISSN: 2038-2529 [Electronic] United States |
PMID | 23389369
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- TNF-Related Apoptosis-Inducing Ligand
- TNFSF10 protein, human
- Sodium Chloride
- Aconitic Acid
- Edetic Acid
- Trypsin
|
Topics |
- Aconitic Acid
- Apoptosis
- Carcinoma, Hepatocellular
(chemistry)
- Cell Adhesion
- Cell Cycle
- Cell Line, Tumor
- Cell Survival
- Edetic Acid
- Flow Cytometry
(methods)
- Humans
- Liver Neoplasms
(chemistry)
- Sodium Chloride
- TNF-Related Apoptosis-Inducing Ligand
(analysis, isolation & purification)
- Trypsin
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