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Article: Effect of cryopreservation on metabolic activities of hepatocytes

TitleEffect of cryopreservation on metabolic activities of hepatocytes
Authors
KeywordsCryopreservation
Cytochrome P-450
Hepatocytes
Metabolic Detoxication, Drug
Issue Date2008
Citation
Chinese Journal of Pharmacology and Toxicology, 2008, v. 22 n. 6, p. 457-462 How to Cite?
AbstractAIM: To study the effect of cryopreservation on metabolic activities and cytochrome P450 (CYP) mRNA expression in hepatocytes and provide support for application of cryopreserved hepatocytes in experimental research. METHODS: Freshly isolated rat hepatocytes were cryopreserved with rate-controlled freezer, and thawed after 1 month. Real-time quantitative PCR was used to detect expressions of CYP1A2, CYP2B1 and CYP3A1 mRNA, and LC-MS/MS was used to measure contents of metabolites of midazolam-1′-hydrxylation (OH-Mid), diclofenac-4′-hydroxylation (OH-Dic) and dextromethorphan-O-demethylation (Dex) in hepatocytes, respectively. RESULTS: There was no significant difference in cell viability between fresh and cryopreserved hepatocytes. The cryopreserved hepatocytes attached and established extensive cell-cell contact, with round and bright nucleus. CYP1 A2 and CYP2B1 mRNA expressions induced by β-naphthoflavone and phénobarbital in cryopreserved hepatocytes were similar to that in the fresh primary cells. However, CYP3A1 mRNA expression did not induced by pregnenolone-16α-carbonitrile in cryopreserved hepatocytes. In cryopreserved hepatocytes, the content of OH-Mid was remained as almost the same as the fresh primary hepatocytes, while contents of OH-Dic decreased approximately as a half, and Dex was double as fresh hepatocytes. CONCLUSION: Cryopreservation exerts different effects on metabolic activities of hepatocytes. To acquire objective and appropriate results, it is necessary to consider the different influence of cryopreservation on cell metabolic activity in drug metabolic research.
Persistent Identifierhttp://hdl.handle.net/10722/92170
ISSN
2015 SCImago Journal Rankings: 0.119
References

 

DC FieldValueLanguage
dc.contributor.authorPan, Qen_HK
dc.contributor.authorChen, Yen_HK
dc.contributor.authorMei, Len_HK
dc.contributor.authorDai, R-Ken_HK
dc.date.accessioned2010-09-17T10:38:08Z-
dc.date.available2010-09-17T10:38:08Z-
dc.date.issued2008en_HK
dc.identifier.citationChinese Journal of Pharmacology and Toxicology, 2008, v. 22 n. 6, p. 457-462en_HK
dc.identifier.issn1000-3002en_HK
dc.identifier.urihttp://hdl.handle.net/10722/92170-
dc.description.abstractAIM: To study the effect of cryopreservation on metabolic activities and cytochrome P450 (CYP) mRNA expression in hepatocytes and provide support for application of cryopreserved hepatocytes in experimental research. METHODS: Freshly isolated rat hepatocytes were cryopreserved with rate-controlled freezer, and thawed after 1 month. Real-time quantitative PCR was used to detect expressions of CYP1A2, CYP2B1 and CYP3A1 mRNA, and LC-MS/MS was used to measure contents of metabolites of midazolam-1′-hydrxylation (OH-Mid), diclofenac-4′-hydroxylation (OH-Dic) and dextromethorphan-O-demethylation (Dex) in hepatocytes, respectively. RESULTS: There was no significant difference in cell viability between fresh and cryopreserved hepatocytes. The cryopreserved hepatocytes attached and established extensive cell-cell contact, with round and bright nucleus. CYP1 A2 and CYP2B1 mRNA expressions induced by β-naphthoflavone and phénobarbital in cryopreserved hepatocytes were similar to that in the fresh primary cells. However, CYP3A1 mRNA expression did not induced by pregnenolone-16α-carbonitrile in cryopreserved hepatocytes. In cryopreserved hepatocytes, the content of OH-Mid was remained as almost the same as the fresh primary hepatocytes, while contents of OH-Dic decreased approximately as a half, and Dex was double as fresh hepatocytes. CONCLUSION: Cryopreservation exerts different effects on metabolic activities of hepatocytes. To acquire objective and appropriate results, it is necessary to consider the different influence of cryopreservation on cell metabolic activity in drug metabolic research.en_HK
dc.languageengen_HK
dc.relation.ispartofChinese Journal of Pharmacology and Toxicologyen_HK
dc.subjectCryopreservationen_HK
dc.subjectCytochrome P-450en_HK
dc.subjectHepatocytesen_HK
dc.subjectMetabolic Detoxication, Drugen_HK
dc.titleEffect of cryopreservation on metabolic activities of hepatocytesen_HK
dc.typeArticleen_HK
dc.identifier.emailChen, Y:ychenc@hkucc.hku.hken_HK
dc.identifier.authorityChen, Y=rp1318en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.3867/j.issn.1000-3002.2008.06.011en_HK
dc.identifier.scopuseid_2-s2.0-58149268050en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-58149268050&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume22en_HK
dc.identifier.issue6en_HK
dc.identifier.spage457en_HK
dc.identifier.epage462en_HK

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