A. Parrella

Intracytoplasmic sperm injection: state of the art in humans

Among infertile couples, 25% involve both male and female factors, while male factor alone accounts for another 25% due to oligo-, astheno-, teratozoospermia, a combination of the three, or even a complete absence of sperm cells in the ejaculate and can lead to a poor prognosis even with the help of assisted reproductive technology (ART). Intracytoplasmic sperm injection (ICSI) has been with us now for a quarter of a century and in spite of the controversy generated since its inception, it remains in the forefront of the techniques utilized in ART. The development of ICSI in 1992 has drastically decreased the impact of male factor, resulting in millions of pregnancies worldwide for couples who, without ICSI, would have had little chance of having their own biological child. This review focuses on the state of the art of ICSI regarding utility of bioassays that evaluate male factor infertility beyond the standard semen analysis and describes the current application and advances in regard to ICSI, particularly the genetic and epigenetic characteristics of spermatozoa and their impact on reproductive outcome.

Sperm DNA fragmentation as treatment guidance for infertile couples

ported only in 2009. Isolation of these cells was replicated by a small number of other groups in mouse, rat and extended to a human model. The objective of this study is to demonstrate the presence of OSC in non-human primates (NHPs), which can generate mature oocytes following transplantation into the ovary. DESIGN: Eggs from ovarian stem cell transplantation were characterized and explored their fertilization potential. MATERIALS AND METHODS: The ovarian cortex was digested with collagenase IV and DNase followed by FACS with the DDX4 antibody. The cells were expanded in culture, transfected with a GFP lentivirus, and transplanted into the remaining ovary of the rhesus monkey. Following transplantation, gonadotropins were used for ovarian hyperstimulation to isolate oocytes. Oocytes transplant derivations were assessed by fluorescence, PCR and nested PCR. True oocyte phenotype with appearance of intact zona pellucida and polar body was observed along with expression of oocyte specific genes. Intracytoplasmic sperm injection was performed with collectedMII oocytes and fertilization and embryo development potential assessed. RESULTS: Here we demonstrate, for the first time, the presence of adult stem cells in the primate ovary which form mature oocytes with fertilization capacity following orthotopic transplantation. Two out of 68 oocytes obtained by follicular aspiration were confirmed OSCs origin, whereas 17 out of 83 from microdissection. A mature oocyte originating from OSCs developed into 64-cells stage embryo. CONCLUSIONS: Stem cells from NHP adult ovaries can be transplanted and give rise to new oocytes that fertilize and develop into an embryo, suggesting that these stem cells could be a novel approach to treating infertility.