title: Sperm chromatin maturity and integrity correlated to zygote development in ICSI program.
abstract: This study aimed to evaluate sperm chromatin maturity and integrity of that injected into good-quality oocytes in an in vitro fertilization-intra cytoplasmic sperm injection (IVF-ICSI) program. A cut-off value of sperm chromatin maturity and integrity was developed as a function of their correlation to the zygote development, i.e., embryo formation and cleavage rate. The study assessed sperm chromatin maturity using aniline blue (AB) staining, whereas toluidine blue (TB) staining was used to assess sperm chromatin integrity. Ejaculates from 59 patients undergoing ICSI and 46 fertile normozoospermic donors for determination of normal values of sperm chromatin status were used in this study. Embryo formation and cleavage rates were observed for the period of 3 days after ICSI. There was a significant difference in the percentage of sperm with mature chromatin between ejaculate from ICSI patients and fertile donor (p=0.020); while there was no significant difference in sperm chromatin integrity of both samples (p=0.120). There was no significant correlation between sperm chromatin maturity and either embryo formation or cleavage rate; as well as sperm chromatin integrity to both parameters of zygote development (p>0.05). Furthermore, we found that the cut-off value of sperm chromatin maturity and integrity of the fertile normozoospermic ejaculates were 87.2% and 80.2%, respectively. Using the cut-offs, we found that low sperm chromatin maturity at the level of <87% correlated significantly with the cleavage rate of the zygote (p=0.022; r=0.371); whereas poor sperm chromatin integrity at the level of <80% correlated with embryo formation (p=0.048; r=0,485). In conclusion, this study showed that poor maturity and integrity of sperm chromatin (AB<87% and TB<80%, respectively), could affect zygote development following ICSI.
  
  
  BACKGROUND
  AB: aniline blue; CMA3: chromomycin A3; ICSI: intra cytoplasmic sperm injection; IVF: in vitro fertilization; PBS: phosphate buffer saline; SPSS: Statistical Package for Social Science; TB: toluidine blue; WHO: World Health Organization.
authors.affiliation: a Department of Medical Biology, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
authors.lastname: Asmarinah
authors.affiliation: b Master Program in Biomedical Sciences, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Syauqy
authors.affiliation: b Master Program in Biomedical Sciences, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
authors.forename: Liya Agustin
authors.initials: LA
authors.lastname: Umar
authors.affiliation: a Department of Medical Biology, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
authors.forename: Silvia Werdhy
authors.initials: SW
authors.lastname: Lestari
authors.affiliation: c Yasmin Fertility Clinic , Cipto Mangunkusumo Hospital , Jakarta , Indonesia.
authors.forename: Eliza
authors.initials: E
authors.lastname: Mansyur
authors.affiliation: c Yasmin Fertility Clinic , Cipto Mangunkusumo Hospital , Jakarta , Indonesia.|d Reproductive Immunoendocrinology Division, Department of Obstetrics and Gynecology, Faculty of Medicine , University of Indonesia , Jakarta , Indonesia.
authors.forename: Andon
authors.initials: A
authors.lastname: Hestiantoro
authors.affiliation: e Department of Urology, Pediatric Urology and Andrology, Faculty of Medicine , University of Giessen , Giessen , Hesse , Germany.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Paradowszka-Dogan
mesh_terms: D000328:Adult; D002843:Chromatin / Q000502:physiology*; D005260:Female; D006801:Humans; D008297:Male; D020554:Sperm Injections, Intracytoplasmic; D013079:Sperm Maturation; D015053:Zygote / Q000254:growth & development*
chemical_list: D002843:Chromatin
keywords: IVF-ICSI; Integrity; maturity; sperm chromatin; zygote development