Assessing the Impact of the Novel Sperm Selection Technique 'Annexin-V Coated Polystyrene Bead Technique' on Mouse Assisted Reproductive Techniques Outcomes: Preliminary Findings

Abstract

ICSI is one of the most commonly used techniques to treat infertility. The sperm selection for the procedure is done 'randomly' by the embryologist according to the motility and morphology parameters which is known not to reflect the potential of a sperm for fertilization, pregnancy and a healthy childbearing. Since the apoptosis rate is higher in sperm cells of infertile patients, it is more likely to choose an apoptotic sperm by the 'random selection method'. We recently introduced a novel sperm selection technique namely 'Annexin-V coated polystrene bead technique'(APB-Tech), for the selection of non-apoptotic sperm cells. The principal of the technique is based on the binding affinity of an apoptotic sperm to 'Annexin-V covered beads' enabling to distinguish a viable and a healthy sperm by light microscopy. The aim of the present study was to observe the effects of this technique on ICSI outcomes in mice. Sibling-oocyte trial was conducted and the outcome measures were compared with the results of traditional sperm selection method. Embryo and blastocyst qualities and blastocyst development rates were significantly increased in APB-Tech group, while the other parameters were not affected. Promising results obtained from the technique reflect its promising potential as a new and powerful tool for sperm selection and thus infertility techniques.

Keywords: APB-Tech; Embryo; ICSI; Infertility; Sperm selection.

Fig. 1

Sperm retrieval procedures are demonstrated A. Testis removal B. Testicular tissue cut into pieces are demonstrated

Fig. 2

Pictures after the sperm suspension is exposed to the APB solution including the beads. Arrows (➨) show bead bounded sperm cells with DNA fragmentation, asterix (★) show unbound sperm cells with intact DNA (X40)

Fig. 3

Sperm selection steps via APB-tech were demonstrated on a microinjection dish for the microinjection (ICSI) procedure

Fig. 4

The ovary and Fallopian tubes are demonstrated after mouse decapitation (A). Ovary and Fallopian tubes are isolated and dissected under a stereo microscope (B). Ampulla region (C) and the contents of the ampulla shed after the rupture of the ampulla region (D). Cumulus-oocyte complexes (COC) are demonstrated (E). Photographs of an oocyte cell surrounded with corona radiata cells are demonstrated (X40)

Fig. 5

Oocyte cells after denudation are demonstrated at different stages of maturity. A mature oocyte in the Metaphase 2 (MII stage) (Polar Body is shown by the arrow) (A), an immature (immature) oocyte in the Metaphase 1 (MI stage) where the polar body is not observed (B), and an immature oocyte in the germinal vesicle (GV stage) shown by the arrow (X40) (C)

Fig. 6

The micromanipulator that the microinjection process is performed (A), the dish in which the oocytes are placed during the microinjection process (B), and the microinjection process (C) are demonstrated (X20)

Fig. 7

A non-fertilized (A) and a fertilized oocyte (B) with 2 pronuclei (arrows) and 2 polar bodies (dashed arrows) are demonstrated (X40)

Fig. 8

Degenerated oocytes (A, B) (cytoplasms are dark colored and retracted) are demonstrated (X20)

Fig. 9

2-cell good quality (A) and poor quality embryo (B) at 24 hours; 4-cell good quality (C) and poor quality embryo (D) at 48 hours (Day 2); 8-cell good quality (E) and poor quality embryo (F) at 72 hours (Day 3) and, compacting embryos (G) and compact embryo (H) at the 96th hour (Day 4); Cavitating embryo (I; J) at 100 hours; Good quality (K) and poor quality blastocyst stage embryos (L) at 120 hours and, a hatching embryo (M) at 130 hours is demonstrated (X40)

Fig. 9

2-cell good quality (A) and poor quality embryo (B) at 24 hours; 4-cell good quality (C) and poor quality embryo (D) at 48 hours (Day 2); 8-cell good quality (E) and poor quality embryo (F) at 72 hours (Day 3) and, compacting embryos (G) and compact embryo (H) at the 96th hour (Day 4); Cavitating embryo (I; J) at 100 hours; Good quality (K) and poor quality blastocyst stage embryos (L) at 120 hours and, a hatching embryo (M) at 130 hours is demonstrated (X40)

Fig. 9

2-cell good quality (A) and poor quality embryo (B) at 24 hours; 4-cell good quality (C) and poor quality embryo (D) at 48 hours (Day 2); 8-cell good quality (E) and poor quality embryo (F) at 72 hours (Day 3) and, compacting embryos (G) and compact embryo (H) at the 96th hour (Day 4); Cavitating embryo (I; J) at 100 hours; Good quality (K) and poor quality blastocyst stage embryos (L) at 120 hours and, a hatching embryo (M) at 130 hours is demonstrated (X40)

Fig. 10

Top quality embryo rates on Day 2, 3 and 5