Influence of electroporation on chicken blastoderm cell viability in vitro.
The aim of this study was to compare two types of devices used for blastoderm cell (BC) transfection: the Nucleofector (Amaxa, Biosystems) and the Multiporator (Eppendorf). To assess the influence of electric current on BCs, different conditions of both nucleofection and electroporation were used. Next, the viability of cells was assessed. The highest number of cells (90.8%) was viable after nucleofection in the G10 program. After transfection in the presence of pmaxGFP, the A23 program was found to be most advantageous. The elecroporation experiment with the Multiporator (Eppendorf) showed a significant influence of osmotic pressure and voltage on BC viability. Namely, in the isoosmolar buffer BC viability was statistically higher (P < or = 0.05) in comparison to the hypoosmolar buffer. The, viability of cells was statistically higher (P < or = 0.05) after application of 25V as compared to 50V. The efficiency of transfection in the presence of EGFP-C 1 after electroporation in 2 pulses, 25V, 500 micros in the isoosmolar buffer was better than in the recommended conditions in the Amaxa Biosystems A23 program.
Wawrzynska
Magdalena
M
Department of Animal Biotechnology, Faculty of Animal Breeding and Biology, University of Technology and Life Sciences, Mazowiecka 28, 85-084 Bydgoszcz, Poland. magda-waw@wp.pl
Bednarczyk
Marek
M
Łakota
Pawel
P
Lubiszewska
Marta
M
D000818:Animals; D001756:Blastoderm / Q000166:cytology*; D002470:Cell Survival / Q000502:physiology; D002645:Chickens; D018274:Electroporation / Q000295:instrumentation* / Q000379:methods; D066298:In Vitro Techniques; D008856:Microscopy, Fluorescence; D014162:Transfection / Q000295:instrumentation* / Q000379:methods