Hamilton De Martin1,2, Marcello S Cocuzza3,4, Bruno C Tiseo1,2, Guilherme J A Wood1,2, Eduardo P Miranda1,2, Pedro A A Monteleone1,5, José Maria Soares1,5, Paulo C Serafini1,5, Miguel Srougi1,2, Edmund C Baracat1,5. 1. Human Reproduction Center, University of Sao Paulo Medical School, Av. Dr. Eneas de Carvalho Aguiar 255, Sao Paulo, 05403-000, Brazil. 2. Division of Urology, University of Sao Paulo Medical School, Sao Paulo, Brazil. 3. Human Reproduction Center, University of Sao Paulo Medical School, Av. Dr. Eneas de Carvalho Aguiar 255, Sao Paulo, 05403-000, Brazil. mcocuzza@uol.com.br. 4. Division of Urology, University of Sao Paulo Medical School, Sao Paulo, Brazil. mcocuzza@uol.com.br. 5. Discipline of Gynecology, University of Sao Paulo Medical School, Sao Paulo, Brazil.
Abstract
PURPOSE: The purpose of this study was to develop a novel one-step ICSI approach to select sperm with better chromatin maturity than the conventional method. METHODS: This was a pilot diagnostic study, which prospectively recruited men during a 6-month period in a University-affiliated infertility centre. Forty consecutive semen samples were provided for analysis. The positive rheotaxis extended drop (PRED) was set up creating a pressure and viscosity gradient. Each semen sample was divided into four aliquots: one aliquot for density gradient centrifugation (DGC), two aliquots for PRED (fresh semen (PRED-FS) and processed semen (PRED-DGC)), and one aliquot as the control (FS). In PRED, a mean of 200 spermatozoa were collected consecutively without selection from the outlet reservoir. The aniline blue assay was used to assess chromatin immaturity. RESULTS: The mean channel length, measured from inlet to outlet, was 32.55 ± 0.86 mm, with a mean width of 1.04 ± 0.21 mm. In 82.5% of cases (33/40), at least 50 spermatozoa were captured between 15 and 30 min. Improved chromatin maturity after the DGC preparation and the PRED approach was observed in all samples. This was reflected by a mean reduction from 28.65 ± 8.97% uncondensed chromatin in the native ejaculates to 17.29 ± 7.72% in DGC and 0.89 ± 1.31% in the PRED approach (P < 0.01). CONCLUSIONS: The PRED method may improve the current ICSI technique by providing it with its own sperm selection process. ICSI would probably become an even more complete technique comprising selection, capture and injection of the male gamete.
PURPOSE: The purpose of this study was to develop a novel one-step ICSI approach to select sperm with better chromatin maturity than the conventional method. METHODS: This was a pilot diagnostic study, which prospectively recruited men during a 6-month period in a University-affiliated infertility centre. Forty consecutive semen samples were provided for analysis. The positive rheotaxis extended drop (PRED) was set up creating a pressure and viscosity gradient. Each semen sample was divided into four aliquots: one aliquot for density gradient centrifugation (DGC), two aliquots for PRED (fresh semen (PRED-FS) and processed semen (PRED-DGC)), and one aliquot as the control (FS). In PRED, a mean of 200 spermatozoa were collected consecutively without selection from the outlet reservoir. The aniline blue assay was used to assess chromatin immaturity. RESULTS: The mean channel length, measured from inlet to outlet, was 32.55 ± 0.86 mm, with a mean width of 1.04 ± 0.21 mm. In 82.5% of cases (33/40), at least 50 spermatozoa were captured between 15 and 30 min. Improved chromatin maturity after the DGC preparation and the PRED approach was observed in all samples. This was reflected by a mean reduction from 28.65 ± 8.97% uncondensed chromatin in the native ejaculates to 17.29 ± 7.72% in DGC and 0.89 ± 1.31% in the PRED approach (P < 0.01). CONCLUSIONS: The PRED method may improve the current ICSI technique by providing it with its own sperm selection process. ICSI would probably become an even more complete technique comprising selection, capture and injection of the male gamete.
Authors: Brenda S Cho; Timothy G Schuster; Xiaoyue Zhu; David Chang; Gary D Smith; Shuichi Takayama Journal: Anal Chem Date: 2003-04-01 Impact factor: 6.986
Authors: Monica Muratori; Nicoletta Tarozzi; Marta Cambi; Luca Boni; Anna Lisa Iorio; Claudia Passaro; Benedetta Luppino; Marco Nadalini; Sara Marchiani; Lara Tamburrino; Gianni Forti; Mario Maggi; Elisabetta Baldi; Andrea Borini Journal: Medicine (Baltimore) Date: 2016-05 Impact factor: 1.889
Authors: Hamilton De Martin; Eduardo P Miranda; Marcello S Cocuzza; Pedro A A Monteleone Journal: J Assist Reprod Genet Date: 2019-10-29 Impact factor: 3.412
Authors: Erica T Y Leung; Cheuk-Lun Lee; Xinyi Tian; Kevin K W Lam; Raymond H W Li; Ernest H Y Ng; William S B Yeung; Philip C N Chiu Journal: Nat Rev Urol Date: 2021-11-05 Impact factor: 14.432