Interleukin-1 alpha is released during transfection of keratinocytes.

Abstract

Keratinocytes are known to produce, store, and release IL-1 alpha and therefore we suspected that the DNA-mediated cell transfection procedure may release the stored IL-1 alpha from keratinocytes into the medium. Using enzyme-linked immunosorbent assay, we determined the IL-1 alpha concentration in culture supernatants during keratinocyte transfection. The following transfection methods were compared: lipofection with lipofectACE and lipofectAMINE (GIBCO), Ca3(PO4)2 co-precipitation, and polybrene-dimethylsulfoxide (DMSO). The supernatants were collected immediately prior to transfection, after 5-h incubation with lipofectin or Ca3(PO4)2, and 24 and 48 h after transfection. In the polybrene-DMSO method, the supernatant was also collected immediately before and after DMSO shock. LipofectAMINE caused the highest release of IL-1 alpha, whereas the lipofectACE and polybrene-DMSO mediated transfection with confluent cells released the least. The other two methods released intermediate levels of IL-1 alpha. Our data indicate that a substantial amount of IL-1 alpha is released during the keratinocyte transfection procedure, which can affect the results of transfection in studies of gene expression.

• Authors: M Komine, I M Freedberg, M Blumenberg
• Journal: The Journal of investigative dermatology (J Invest Dermatol) Vol. 103 Issue 4 Pg. 580-2 (Oct 1994) ISSN: 0022-202X [Print] United States
• PMID: 7930685 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Cation Exchange Resins
• Indicators and Reagents
• Interleukin-1
• Lipids
• Lipofectamine
• Chloramphenicol O-Acetyltransferase
• beta-Galactosidase
• Dimethyl Sulfoxide

Topics

• Cation Exchange Resins (pharmacology)
• Cell Death (drug effects)
• Chloramphenicol O-Acetyltransferase (metabolism)
• Dimethyl Sulfoxide (pharmacology)
• Humans
• Indicators and Reagents (pharmacology)
• Infant, Newborn
• Interleukin-1 (metabolism)
• Keratinocytes (drug effects, metabolism, physiology)
• Lipids (pharmacology)
• Male
• Transfection
• beta-Galactosidase (metabolism)