Preimplantation development of mouse oocytes following intracytoplasmic injection with preserved GFP spermatozoa at - 80 degrees C

Abstract

The purpose of this study was to produce the GFP embryo from preserved GFP spermatozoa with ICR mouse background at -80oC (GFP spermatozoa) via Intracytoplasmic Sperm Injection (ICSI) technique. Mouse spermatozoa were collected from two cauda epididymis of mature mice. All experiments were performed with mature oocytes which were collected from adult ICR female mouse after superovulation inducing. After GFP spermatozoa were collected and transported from Japan to Viet Nam, it was stored in HEPES-mCZB medium containing NaCl and keep it at -80oC until use. ICSI technique was performed with fresh ICR spermatozoa for control experiment to create Fresh sperm embryos, and GFP spermatozoa in order to generate GFP embryos. As a result, the preimplantation development of GFP embryos and Fresh sperm embryos were 21.43% and 26.19%, respectively. Moreover, the total number of cell from the early GFP blastocyst and the early Fresh sperm blastocyst embryos were 30.75 ± 2.50 and 29.25 ± 5.32, respectively. The results indicated there was no significant difference between the development of GFP embryos and Fresh sperm embryos via ICSI after use Paired T–test to compare the difference between two samples. In general conclusion, the oocytes were injected with GFP spermatozoa preserved at -80oC in HEPES-mCZB medium containing NaCl after collected at Japan and then transported to Viet Nam, which could develop to the blastocyst stage embryos.