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Novel distance-progesterone-combined selection approach improves human sperm quality.

ABSTRACT: BACKGROUND:Sperm selection is essential for the health of offspring conceived via assistive reproductive technology (ART). Various methods of sperm preparation for in vitro fertilization and intracytoplasmic sperm injection have been developed to acquire sperm with better quality and to avoid potential genetic disorders. However, current sperm processing and selection techniques bypass the natural selection that occurs during fertilization in vivo. The aim of this study was to present a novel distance-progesterone-combined selection approach with an original device based on the human female reproductive tract, and to report on its effectiveness based on sperm progressive motility, as well as chemotaxis. METHODS:A novel device with long distance channels which mimicked the female human reproductive system was designed and fabricated. This ready-to-be-used device was developed using a progesterone gradient and human tube fluid media. Sperm swam for 150 min in the device under conditions of 37 °C air temperature with 5% CO2 after separation from seminal plasma via discontinuous Percoll gradient treatment. The selected sperm were assessed for normal morphology using Diff-Quik staining. A chromatin diffusion assay assessed sperm for DNA fragments and apoptosis was assessed using annexin V-fluorescein isothiocyanate/propidium iodide fluorescent staining. RESULTS:Our distance-progesterone-combined sperm selection method was successfully established. After sperm were selected, the percentage of sperm with normal morphology increased (before vs. after selection, 11.2?±?1.3% vs. 40.3?±?6.6%, P?=?0.000), the percentage of sperm with DNA fragmentation decreased (before vs. after selection, 15.4?±?4.0% vs. 6.8?±?3.3%, P?=?0.001), and the percentage of sperm with apoptosis did not change significantly. CONCLUSIONS:Our newly-developed method is capable of successfully selecting sperm of high quality. The method will be benefit clinical ART practice as it can reduce sperm-related genetic risks.

SUBMITTER: Li K

PROVIDER: S-EPMC6053761 | biostudies-literature | 2018 Jul

REPOSITORIES: biostudies-literature

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Novel distance-progesterone-combined selection approach improves human sperm quality.

Li Kun K Li Rui R Ni Ya Y Sun Peibei P Liu Ye Y Zhang Dan D Huang Hefeng H

Journal of translational medicine 20180720 1

<h4>Background</h4>Sperm selection is essential for the health of offspring conceived via assistive reproductive technology (ART). Various methods of sperm preparation for in vitro fertilization and intracytoplasmic sperm injection have been developed to acquire sperm with better quality and to avoid potential genetic disorders. However, current sperm processing and selection techniques bypass the natural selection that occurs during fertilization in vivo. The aim of this study was to present a ...[more]

PMID: 30029659

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Project description:Study questionCan we make the comet assay (single-cell gel electrophoresis) for human sperm a more accurate and informative high throughput assay?Summary answerWe developed a standardized automated high throughput comet (HT-COMET) assay for human sperm that improves its accuracy and efficiency, and could be of prognostic value to patients in the fertility clinic.What is known alreadyThe comet assay involves the collection of data on sperm DNA damage at the level of the single cell, allowing the use of samples from severe oligozoospermic patients. However, this makes comet scoring a low throughput procedure that renders large cohort analyses tedious. Furthermore, the comet assay comes with an inherent vulnerability to variability. Our objective is to develop an automated high throughput comet assay for human sperm that will increase both its accuracy and efficiency.Study design, size, durationThe study comprised two distinct components: a HT-COMET technical optimization section based on control versus DNAse treatment analyses ( ITALIC! n = 3-5), and a cross-sectional study on 123 men presenting to a reproductive center with sperm concentrations categorized as severe oligozoospermia, oligozoospermia or normozoospermia.Participants/materials, setting, methodsSperm chromatin quality was measured using the comet assay: on classic 2-well slides for software comparison; on 96-well slides for HT-COMET optimization; after exposure to various concentrations of a damage-inducing agent, DNAse, using HT-COMET; on 123 subjects with different sperm concentrations using HT-COMET. Data from the 123 subjects were correlated to classic semen quality parameters and plotted as single-cell data in individual DNA damage profiles.Main results and the role of chanceWe have developed a standard automated HT-COMET procedure for human sperm. It includes automated scoring of comets by a fully integrated high content screening setup that compares well with the most commonly used semi-manual analysis software. Using this method, a cross-sectional study on 123 men showed no significant correlation between sperm concentration and sperm DNA damage, confirming the existence of hidden chromatin damage in men with apparently normal semen characteristics, and a significant correlation between percentage DNA in the tail and percentage of progressively motile spermatozoa. Finally, the use of DNA damage profiles helped to distinguish subjects between and within sperm concentration categories, and allowed a determination of the proportion of highly damaged cells.Limitations, reasons for cautionThe main limitations of the HT-COMET are the high, yet indispensable, investment in an automated liquid handling system and heating block to ensure accuracy, and the availability of an automated plate reading microscope and analysis software.Wider implications of the findingsThis standardized HT-COMET assay offers many advantages, including higher accuracy and evenness due to automation of sensitive steps, a 14.4-fold increase in sample analysis capacity, and an imaging and scoring time of 1 min/well. Overall, HT-COMET offers a decrease in total experimental time of more than 90%. Hence, this assay constitutes a more efficient option to assess sperm chromatin quality, paves the way to using this assay to screen large cohorts, and holds prognostic value for infertile patients.Study funding/competing interestsFunded by the CIHR Institute of Human Development, Child and Youth Health (IHDCYH; RHF 100625). O.A. is a fellow supported by the Fonds de la Recherche du Québec - Santé (FRQS) and the CIHR Training Program in Reproduction, Early Development, and the Impact on Health (REDIH). B.R. is a James McGill Professor. The authors declare no conflicts of interest.

Dataset Information

Novel distance-progesterone-combined selection approach improves human sperm quality.

Publications

Novel distance-progesterone-combined selection approach improves human sperm quality.

Similar Datasets

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