Dimethyl sulphoxide and electrolyte-free medium improve exogenous DNA uptake in mouse sperm and subsequently gene expression in the embryo
Autor: | Soleiman Kurd, Mohammad Salehi, Vahid Jajarmi, Sara Hosseini, Fardin Fathi |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Male Cell Survival medicine.medical_treatment Green Fluorescent Proteins Mice Inbred Strains Mice Transgenic Fertilization in Vitro Andrology 03 medical and health sciences Sperm-mediated gene transfer Electrolytes 0302 clinical medicine Human fertilization medicine Animals Humans Dimethyl Sulfoxide Blastocyst reproductive and urinary physiology Sperm motility 030219 obstetrics & reproductive medicine In vitro fertilisation urogenital system Chemistry Gene Expression Regulation Developmental Embryo Cell Biology DNA Sperm Spermatozoa Culture Media 030104 developmental biology medicine.anatomical_structure Sperm Motility Exogenous DNA Female Developmental Biology |
Zdroj: | Zygote (Cambridge, England). 26(5) |
ISSN: | 1469-8730 |
Popis: | SummaryOne of the methods to generate transgenic animals is called sperm-mediated gene transfer (SMGT). Mature sperm cells can take up exogenous DNA molecules intrinsically and transfer them into the oocyte during fertilization. This study assessed the effect of dimethyl sulfoxide (DMSO) and electrolyte-free medium (EFM) on DNA uptake (EGFP–N1plasmid) in mouse sperm. Sperms cells cultured in human tubular fluid (HTF) without any treatment were considered as the control group. Sperms cells that were incubated in EFM and HTF with DNA/DMSO at 4°C were classified into EFM and HTF groups. Sperm motility and viability were assessed following treatment. In vitro fertilization (IVF) with sperm in all groups was performed. Fertilization, embryo development and GFP-positive blastocyst rates were analyzed and compared. The result showed that sperm motility and viability in EFM were better than those in the HTF group. The rate of development to reach the blastocyst stage and GFP-positive blastocysts was significantly higher in the EFM group compared with the HTF group (P |
Databáze: | OpenAIRE |
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