Morphokinetic evaluation of embryos generated from vitrified oocytes maintaining the meiotic spindle

Vitrification is a technique for preservation of human oocytes. There is still a lack of basic research about the possible effects of vitrification on subsequent embryos following oocyte vitrification. The purpose of this study was to evaluate the embryo morphokinetic parameters formed after fertilization of vitrified-warmed oocytes, where an intact meiotic spindle (MS) was observed pre- and post-cryopreservation. Matured oocytes after in vitro maturation were collected and MS evaluation was performed. The oocytes with MS were divided into two groups: fresh and post vitrification. After intra-cytoplasmic sperm injection, the oocytes were cultured in time lapse monitoring (TLM) and time of second polar body extrusion (SPBE), pronuclei appearance (tPNA), pronuclei fading (tPNF), formation of two to eight cells (t2 to t8), and irregular cleavage events [direct cleavage (DC), reverse cleavage (RC)] and vacuolation were assessed. The fertilization rate was not significantly different between the groups, although the rate of abnormal fertilization was higher in vitrification group compared with fresh group (23.5% VS 7.7%). Analysis of the TLM showed a significant delay in time points, including SPBE, tPNA, tPNF, t 2-cells cleavage in vitrification group (p = 0.02, p = 0.00, p = 0.002, P = 0.00, P = 0.01, respectively). In addition, t3 and t4 time points tended to be delayed in vitrification group (p = 0.05). Moreover, the higher level of DC, RC and vacuolation were noticed in the vitrification group (P˂0.05). In conclusion, despite MS maintenance after warming, TLM evaluation showed both a delay and abnormal cleavage patterns in generated embryos.