Establishing the normal range of sperm DNA fragmentation index (% DFI) for rhesus macaques

Fernanda C. Burch; Philberta Y. Leung; Eric McDonald; Jared Jensen; Emily Mishler; Nadine Piekarski; Camilla M. Mendes; Andrew Sylwester; Carol B. Hanna

doi:10.1038/s41598-023-46928-w

Establishing the normal range of sperm DNA fragmentation index (% DFI) for rhesus macaques

Fernanda C. Burch, Philberta Y. Leung, Eric McDonald, Jared Jensen, Emily Mishler, Nadine Piekarski, Camilla M. Mendes, Andrew Sylwester, Carol B. Hanna

Research output: Contribution to journal › Article › peer-review

Abstract

The Sperm Chromatin Structure Assay (SCSA) is a robust test with high repeatability and precision. It is a clinically accepted assay that defines risk for infertility in men by measuring the degree of DNA fragmentation (% DFI) in sperm. The objective of this study was to adapt and validate the SCSA for rhesus macaques (Macaca mulatta) and establish a range for % DFI in fertile males. Sperm samples from two different males were used to produce a % DFI validation curve before establishing a range using additional samples from n = 11 males. Sperm labeled with acridine orange were analyzed by flow cytometry to measure green fluorescence (native or intact DNA) and red fluorescence (fragmented DNA). Data were exported to FlowJo software to determine the % DFI for each sample. DNA fragmentation ranged from 0.1 to 2.4% DFI, with a mean ± SD = 1.1 ± 0.7% DFI (validation curve optimized to R² > 0.95). In conclusion, we were able to successfully validate the SCSA in our institution and establish the first normal range for sperm DNA fragmentation in rhesus macaques. Our study provides a quantitative baseline for future evaluations to assess macaque fertility through the SCSA test.

Original language	English (US)
Article number	20016
Journal	Scientific Reports
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41598-023-46928-w
State	Published - Dec 2023

ASJC Scopus subject areas

General

Access to Document

10.1038/s41598-023-46928-w

Cite this

@article{01b549ae3e1642c39d8c4c44d5609181,

title = "Establishing the normal range of sperm DNA fragmentation index (% DFI) for rhesus macaques",

abstract = "The Sperm Chromatin Structure Assay (SCSA) is a robust test with high repeatability and precision. It is a clinically accepted assay that defines risk for infertility in men by measuring the degree of DNA fragmentation (% DFI) in sperm. The objective of this study was to adapt and validate the SCSA for rhesus macaques (Macaca mulatta) and establish a range for % DFI in fertile males. Sperm samples from two different males were used to produce a % DFI validation curve before establishing a range using additional samples from n = 11 males. Sperm labeled with acridine orange were analyzed by flow cytometry to measure green fluorescence (native or intact DNA) and red fluorescence (fragmented DNA). Data were exported to FlowJo software to determine the % DFI for each sample. DNA fragmentation ranged from 0.1 to 2.4% DFI, with a mean ± SD = 1.1 ± 0.7% DFI (validation curve optimized to R2 > 0.95). In conclusion, we were able to successfully validate the SCSA in our institution and establish the first normal range for sperm DNA fragmentation in rhesus macaques. Our study provides a quantitative baseline for future evaluations to assess macaque fertility through the SCSA test.",

author = "Burch, {Fernanda C.} and Leung, {Philberta Y.} and Eric McDonald and Jared Jensen and Emily Mishler and Nadine Piekarski and Mendes, {Camilla M.} and Andrew Sylwester and Hanna, {Carol B.}",

note = "Publisher Copyright: {\textcopyright} 2023, The Author(s).",

year = "2023",

month = dec,

doi = "10.1038/s41598-023-46928-w",

language = "English (US)",

volume = "13",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

TY - JOUR

T1 - Establishing the normal range of sperm DNA fragmentation index (% DFI) for rhesus macaques

AU - Burch, Fernanda C.

AU - Leung, Philberta Y.

AU - McDonald, Eric

AU - Jensen, Jared

AU - Mishler, Emily

AU - Piekarski, Nadine

AU - Mendes, Camilla M.

AU - Sylwester, Andrew

AU - Hanna, Carol B.

PY - 2023/12

Y1 - 2023/12

N2 - The Sperm Chromatin Structure Assay (SCSA) is a robust test with high repeatability and precision. It is a clinically accepted assay that defines risk for infertility in men by measuring the degree of DNA fragmentation (% DFI) in sperm. The objective of this study was to adapt and validate the SCSA for rhesus macaques (Macaca mulatta) and establish a range for % DFI in fertile males. Sperm samples from two different males were used to produce a % DFI validation curve before establishing a range using additional samples from n = 11 males. Sperm labeled with acridine orange were analyzed by flow cytometry to measure green fluorescence (native or intact DNA) and red fluorescence (fragmented DNA). Data were exported to FlowJo software to determine the % DFI for each sample. DNA fragmentation ranged from 0.1 to 2.4% DFI, with a mean ± SD = 1.1 ± 0.7% DFI (validation curve optimized to R2 > 0.95). In conclusion, we were able to successfully validate the SCSA in our institution and establish the first normal range for sperm DNA fragmentation in rhesus macaques. Our study provides a quantitative baseline for future evaluations to assess macaque fertility through the SCSA test.

AB - The Sperm Chromatin Structure Assay (SCSA) is a robust test with high repeatability and precision. It is a clinically accepted assay that defines risk for infertility in men by measuring the degree of DNA fragmentation (% DFI) in sperm. The objective of this study was to adapt and validate the SCSA for rhesus macaques (Macaca mulatta) and establish a range for % DFI in fertile males. Sperm samples from two different males were used to produce a % DFI validation curve before establishing a range using additional samples from n = 11 males. Sperm labeled with acridine orange were analyzed by flow cytometry to measure green fluorescence (native or intact DNA) and red fluorescence (fragmented DNA). Data were exported to FlowJo software to determine the % DFI for each sample. DNA fragmentation ranged from 0.1 to 2.4% DFI, with a mean ± SD = 1.1 ± 0.7% DFI (validation curve optimized to R2 > 0.95). In conclusion, we were able to successfully validate the SCSA in our institution and establish the first normal range for sperm DNA fragmentation in rhesus macaques. Our study provides a quantitative baseline for future evaluations to assess macaque fertility through the SCSA test.

UR - http://www.scopus.com/inward/record.url?scp=85176964677&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85176964677&partnerID=8YFLogxK

U2 - 10.1038/s41598-023-46928-w

DO - 10.1038/s41598-023-46928-w

M3 - Article

C2 - 37974016

AN - SCOPUS:85176964677

SN - 2045-2322

VL - 13

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 20016

ER -

Establishing the normal range of sperm DNA fragmentation index (% DFI) for rhesus macaques

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this