Efficient adenoviral transduction of 3T3-F442A preadipocytes without affecting adipocyte differentiation.
The preadipocyte cell lines 3T3-L1 and 3T3-F442A are widely used to study the cellular mechanisms of preadipocyte differentiation and mature adipocyte functions. However, transfection with naked DNA is inefficient in these cell lines. Adenoviral gene transfer is a powerful technique to induce high levels of transgene expression. After failing to obtain 3T3-F442A stable transfectants, we studied different techniques designed to enhance the efficiency of adenoviral transduction in fat cells. First, we compared the effects of two agents known to significantly enhance adenoviral transgene transduction, namely the cationic lipid lipofectamine and the cationic polymer polylysine. We show here that lipofectamine-assisted adenoviral transduction was more efficient in 3T3-F442A than in 3T3-L1 preadipocytes at all tested multiplicity of infection. Lipofectamine, and more efficiently polylysine, yielded high and sustained levels of adenoviral transgene expression in 3T3-F442A preadipocytes. Adenoviral transgene expression was maintained throughout the differentiation process. Furthermore, the two agents also efficiently enhanced adenoviral transduction in mature 3T3-F442A adipocytes. Interestingly, neither protocol affected the differentiation process, morphological features or protein expression of mature adipocytes. These approaches could be of interest to study fat cell differentiation and the functions of mature adipocytes.
Inserm U.680, Faculté de médecine Saint-Antoine, Université Pierre et Marie Curie, 27, rue Chaligny, 75571 Paris cedex 12, France.
Véronique
V
Béréziat
Sylviane
S
Moritz
Bernard
B
Klonjkowski
Aurélie
A
Decaudain
Martine
M
Auclair
Marc
M
Eloit
Jacqueline
J
Capeau
Corinne
C
Vigouroux
D041721:3T3-L1 Cells; D000256:Adenoviridae / Q000235:genetics; D017667:Adipocytes / Q000166:cytology* / Q000187:drug effects; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D008055:Lipids / Q000494:pharmacology; D051379:Mice; D011107:Polylysine / Q000494:pharmacology; D014161:Transduction, Genetic / Q000379:methods*
D008055:Lipids; C086724:Lipofectamine; D011107:Polylysine