Single-blind clinical trial: Sperm selection based on capacity to pass through cumulus oophorous column improves ICSI outcomes.

Background: Sperm selection procedures for future strategies that aim to select normal spermatozoa with intact DNA to improve intracytoplasmic sperm injection (ICSI) outcomes are in early developing stage.

Objectives: The objective is to find out whether the sperm selection procedure based on the ability of spermatozoa to traverse the cumulus cells could improve clinical outcomes of ICSI technique in infertile couples with male factor etiology.

Materials And Methods: For this single-blind clinical trial, mature metaphase II oocytes were retrieved from 150 couples with male factor infertility, male age lower than 45 years and female age under 38 years.

Preparation, Assembly, and Transduction of Transgenic Elements Using Mosaic Analysis with Dual Recombinases (MADR).

This protocol focuses on the cloning and stable integration of sequences of interest by the use of a mosaic analysis with dual recombinases (MADR) plasmid that includes fusion proteins or independent proteins under the control of 2A peptide or IRES elements. Additionally, we describe how to generate a neural stem cell culture from Gt(ROSA)26SortJ mice, and validate the MADR plasmids and by neonatal mouse brain electroporation. This protocol can be generalized to analyze any transgenic element using MADR technology.

Rapid Generation of Somatic Mouse Mosaics with Locus-Specific, Stably Integrated Transgenic Elements.

In situ transgenesis methods such as viruses and electroporation can rapidly create somatic transgenic mice but lack control over copy number, zygosity, and locus specificity. Here we establish mosaic analysis by dual recombinase-mediated cassette exchange (MADR), which permits stable labeling of mutant cells expressing transgenic elements from precisely defined chromosomal loci. We provide a toolkit of MADR elements for combination labeling, inducible and reversible transgene manipulation, VCre recombinase expression, and transgenesis of human cells.