Voltages up to 600V did not affect cryopreserved bovine spermatozoa on capillary-type electroporation

Efficiency and cell viability implications using tip type electroporation in zebrafish sperm cells

“…It has been previously shown that movement variables such as VAP and VSL are strongly correlated with fertilization rates in Anthocidaris crassispina [42]. These results are in agreement with our previous study in bovine sperm cells using tip type electroporation [11]. In a similar way, at high voltages, there was an increased loss of integrity related to damaged plasma membrane, while at low voltages, cell integrity was slight reduced in comparison to non-electroporated spermatozoa.…”

“…Our experiments suggest that capillary electroporation does not affect mitochondrial function. These results corroborate with our previous results using tip type electroporation in bovine sperm, in which reduced motility was caused by electroporation conditions rather than loss of mitochondrial functionality [11].…”

“…Cells with intact membranes were differentiated from permeable cells by their ability to exclude propidium iodide (PI) that easily penetrate dead or damaged cells. The sperm were classified as intact (CDFA + /PI−) and permeable (CDFA + /PI + ; CDFA -/ PI + ; CDFA -/PI−) [11]. The rate was calculated from the number of not-damaged sperm cells with functional membrane/number of sperm cells positive for H 33342, this number was multiplied by 100 to express the percentage of spermatozoa for each category and the result were expressed as percentage of cells with intact membranes.…”

“…In this configuration, the negative effects of the conventional electroporation system, such as change in pH and generation of metal ions, harmful to the cells, are quite reduced, leading to a safer and more efficient transfection. Recently, we successful demonstrated the electrotransfection in cryopreserved bovine spermatozoa through a new protocol using the tip type electroporation [11]. However, no other studies using tip type electroporation for sperm cells have been conducted.…”

“…Despite increased DNA uptake by sperm cells, some problems are associated with the conventional cuvette type electroporation, such as low transfection rate, mosaic gene expression, and negative effects on cell viability [ 9 ]. Capillary electroporation (tip-type electroporation) was first described in 2008 [ 10 ] and since that, it has been consolidated as a new electrotransfection method capable to provide high transfection efficiency on a wide range of mammalian cells with high survival rates of treated cells [ 10 , 11 ]. The mechanical design of tip type electroporation leads to rapid cell delivery by pipette, improving the electroporation efficiency.…”