PMID- 30210017
OWN - NLM
STAT- MEDLINE
DCOM- 20181211
LR  - 20181211
IS  - 1469-8730 (Electronic)
IS  - 0967-1994 (Linking)
VI  - 26
IP  - 4
DP  - 2018 Aug
TI  - Comparison between electroporation and polyfection in pig sperm: efficiency and 
      cell viability implications.
PG  - 286-293
LID - 10.1017/S0967199418000205 [doi]
AB  - SummaryThe aim of this study was to optimize protocols for electroporation (EP) 
      and polyfection (PLF) using polyethyleneimine (PEI) for pig sperm transfection 
      and to determine which method was the most efficient. For EP standardization, 
      different voltages, amounts and times of electric pulses were tested using 
      propidium iodide (PI) as reporter. For PLF standardization, different 
      concentrations of fluorescein isothiocyanate (FITC)-labelled PEI (PEI/FITC) were 
      incubated with sperm for different periods of time. Flow cytometry was performed 
      to evaluate the best protocol in terms of cell viability, including cytoplasmic 
      membrane, acrosome, chromatin integrities and mitochondrial potential using the 
      FITC probe, PI, acridine orange (AO) and JC1. Transfections with the plasmid 
      pmhyGENIE-5 were carried out under optimum conditions for each procedure (EP: 500 
      volts, 500 mus and two pulses; PLF: PEI 0.5 mg/ml and incubation time 10 min). 
      Transfection efficacy was assessed by fluorescence in situ hybridization (FISH). 
      A lower transfection rate was observed for sperm in the control group (17.8%) 
      compared with EP (36.7%), with PLF (76.8%) being the most efficient. These 
      results suggest that the EP and PEI could be an efficient and low cost 
      transfection method for swine sperm. Notably, treated cells showed higher 
      plasmatic the membrane damage (PMD) and/or acrosome damage (AD) indexes, 
      therefore the combination of this procedure with biotechniques that facilitate 
      fecundation (i.e. in vitro fertilization or intracytoplasmic sperm injection) or 
      even inclusion of antioxidant or anti-apoptotic drugs to improve spermatozoa 
      viability would be important.
FAU - da Silva, Zigomar
AU  - da Silva Z
AUID- ORCID: 0000-0002-4356-8910
AD  - 2Santa Catarina State University.College of Agriculture and Veterinary - CAV,Av. 
      Luiz de Camoes,2090,Conta Dinheiro,CEP: 88.520-000,Lages City,SC,Brazil.
FAU - de Souza, Andressa Pereira
AU  - de Souza AP
AD  - 2Santa Catarina State University.College of Agriculture and Veterinary - CAV,Av. 
      Luiz de Camoes,2090,Conta Dinheiro,CEP: 88.520-000,Lages City,SC,Brazil.
FAU - Pandolfi, Jose Rodrigo Claudio
AU  - Pandolfi JRC
AD  - 1Brazilian Agricultural Research Corporation,Embrapa Swine and Poultry,13 BR-153, 
      Km 110,Tamandua District,Post Box: 321 CEP: 89715-899,Concordia, SC.Brazil.
FAU - da Fonseca, Francisco Noe
AU  - da Fonseca FN
AD  - 1Brazilian Agricultural Research Corporation,Embrapa Swine and Poultry,13 BR-153, 
      Km 110,Tamandua District,Post Box: 321 CEP: 89715-899,Concordia, SC.Brazil.
FAU - da Veiga Lima-Rosa, Carlos Andre
AU  - da Veiga Lima-Rosa CA
AD  - 4Santa Catarina State University,College of Southern Region - Ceres,Street Cel. 
      Fernandes Martins,270,Progresso,CEP: 88.790-000,Laguna City,SC,Brazil.
FAU - Marques, Mariana Groke
AU  - Marques MG
AD  - 1Brazilian Agricultural Research Corporation,Embrapa Swine and Poultry,13 BR-153, 
      Km 110,Tamandua District,Post Box: 321 CEP: 89715-899,Concordia, SC.Brazil.
LA  - eng
PT  - Comparative Study
PT  - Journal Article
DEP - 20180913
PL  - England
TA  - Zygote
JT  - Zygote (Cambridge, England)
JID - 9309124
RN  - 9002-98-6 (Polyethyleneimine)
SB  - IM
MH  - Animals
MH  - Cell Survival
MH  - Electroporation/*veterinary
MH  - Fertilization in Vitro
MH  - Male
MH  - Polyethyleneimine/*chemistry
MH  - Semen Preservation/*veterinary
MH  - Sperm Motility
MH  - Spermatozoa/*cytology/physiology
MH  - Swine
MH  - Transfection/*veterinary
OTO - NOTNLM
OT  - Electroporation
OT  - Flow cytometry
OT  - Pig
OT  - Polyethyleneimine
OT  - Sperm viability
EDAT- 2018/09/14 06:00
MHDA- 2018/12/12 06:00
CRDT- 2018/09/14 06:00
PHST- 2018/09/14 06:00 [pubmed]
PHST- 2018/12/12 06:00 [medline]
PHST- 2018/09/14 06:00 [entrez]
AID - S0967199418000205 [pii]
AID - 10.1017/S0967199418000205 [doi]
PST - ppublish
SO  - Zygote. 2018 Aug;26(4):286-293. doi: 10.1017/S0967199418000205. Epub 2018 Sep 13.