Low sample volume causes differentiation in human rhabdomyosarcoma cell line RD subjected to electroporation.
Gene transfection has been accomplished with a variety of techniques such as DEAE dextran, calcium phosphate coprecipitation, protoplast fusion, liposomes, microinjection and recombinant bacteriophages. However, transfection by electroporation, consisting of the reversible permeabilization of cell membranes after exposure to a pulsed electric field, has been shown to be the most rapid, simple and efficient method for the stable incorporation of genes in different cell lines. We studied rhabdomyosarcoma cells subjected to electroporation in two different vol. [400 microliters (group 1) and 150 microliters (group 2] of 140 mM NaCl/15 mM Hepes buffer, pH 7.2) and evaluated the effects of electroporation volume on growth and differentiation. Low sample volumes induced a terminal process of morphological and ultrastructural myogenic differentiation in rhabdomyosarcoma cells, which concluded with cell death. Our results suggest that in electroporation low sample vol. of rhabdomyosarcoma cells induced morphological and phenotypic differentiation, with increased expression of desmin, alpha-actinin and tropomyosin.
Aránega
A
A
Department of Morphological Sciences, School of Medicine, University of Granada, Spain.
Marchal
J A
JA
Melguizo
C
C
Prados
J
J
Aránega
A E
AE
Vélez
C
C
Fernández
J E
JE
Arena
N
N
Alvarez
L
L
D000185:Actinin / Q000302:isolation & purification; D000943:Antigens, Differentiation; D002452:Cell Count; D002454:Cell Differentiation; D002460:Cell Line; D003893:Desmin / Q000302:isolation & purification; D018274:Electroporation; D006801:Humans; D024510:Muscle Development; D018233:Rhabdomyosarcoma, Embryonal / Q000648:ultrastructure*; D014335:Tropomyosin / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D000943:Antigens, Differentiation; D003893:Desmin; D014335:Tropomyosin; D000185:Actinin