Quick recovery and characterization of cell-free DNA in seminal plasma of normozoospermia and azoospermia: implications for non-invasive genetic utilities

Li, Honggang; Huang, Shiyun; Zhou, Hui; Liao, Aihua; Xiong, Chengliang

doi:10.1038/aja.2009.65

Cited by 52 publications

(62 citation statements)

References 18 publications

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“…(4) NGS (Next-generation sequencing) [3] One important aspect of isolated cfDNA is size distribution/fragmentation of it. It can be accessed with electrophoresis.…”

Section: Discussionmentioning

confidence: 99%

“…spermatocytes and spermatids [2]) which results in high concentrations of cfDNA in sperm [3] with heterogenic size molecules [2]. Cell-free nucleic acids (cfDNA and RNA) are released by dying cells, actively secreted by living cells and may have biological role in the organism [3].…”

Section: Biological Aspects Of Cfdnamentioning

confidence: 99%

“…Data: concentration of cfDNA in seminal plasma of normozoospermia: 1.34 ± 0.65 μg mL −1 , with ranges from 0.51 to 2.73 μg mL −1 [3].…”

Section: Biological Aspects Of Cfdnamentioning

confidence: 99%

“…Other applications can longitudinally monitor disease progression and response to treatment [3]. In different studies cell-free DNA was detected in all semen specimens [2].…”

Section: Clinical Applicationmentioning

confidence: 99%

“…In different studies cell-free DNA was detected in all semen specimens [2]. Isolated cfDNA from semen represents easily accessible and non-invasive source of genetic material for further analysis [3].…”

Section: Clinical Applicationmentioning

confidence: 99%

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Isolation of Cell-Free DNA from Seminal Fluid

Draškovič¹

2017

JPP

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Cell-free DNA (cfDNA) is small short double stranded molecule that is also found in seminal fluid. It is a product of apoptotic cells in different developmental stages during spermatogenesis. To final concentration of total cfDNA in semen contributes also cfDNA secreted from living cells and cfDNA that is result of different diseases e.g. prostate cancer or infertility. Amended concentration (high or low) can be connected to prostate cancer or male infertility and can represent important non-invasive diagnostic biomarker for detection and prognosis of these pathological conditions. In this paper, I will discuss different approaches for isolation of cfDNA from seminal fluid, which includes selection of the samples, separation, isolation, extraction, purification and analysis. Today's most popular approach for isolation is the use of commercial kits based on selective binding and elution on silica-membrane technology, magnetic-bead technology or extraction with organic solvents and salting out procedure. Furthermore I will present that I tried to isolate cfDNA from semen with QIAamp DNA Mini Kit to confirm the presence of cell-free DNA in our samples. In the end I will describe problems we are facing during cfDNA measurement which are mainly associated with low concentration of cfDNA in samples.

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“…(4) NGS (Next-generation sequencing) [3] One important aspect of isolated cfDNA is size distribution/fragmentation of it. It can be accessed with electrophoresis.…”

Section: Discussionmentioning

confidence: 99%

Section: Biological Aspects Of Cfdnamentioning

confidence: 99%

“…Data: concentration of cfDNA in seminal plasma of normozoospermia: 1.34 ± 0.65 μg mL −1 , with ranges from 0.51 to 2.73 μg mL −1 [3].…”

Section: Biological Aspects Of Cfdnamentioning

confidence: 99%

“…Other applications can longitudinally monitor disease progression and response to treatment [3]. In different studies cell-free DNA was detected in all semen specimens [2].…”

Section: Clinical Applicationmentioning

confidence: 99%

Section: Clinical Applicationmentioning

confidence: 99%

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Isolation of Cell-Free DNA from Seminal Fluid

Draškovič¹

2017

JPP

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Recovery of cell‐free mRNA and microRNA from human semen based on their physical nature

Wang

Lv²,

Guo

et al. 2014

Biotech and App Biochem

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Cell-free seminal mRNA (cfs-mRNA) and microRNA (cfs-miRNA) have been found in human ejaculate and reported as promising noninvasive biomarkers for disorders of male reproductive organs and forensic identification. However, seminal plasma is particularly challenging for RNA extraction due to its complicated composition and high content of protein, DNA, and polysaccharide. Here, we report a novel, simple, and reliable method for the isolation of cfs-mRNA and cfs-miRNA from human semen based on our previous findings of their physical nature. Seminal microvesicles (0.1-0.5 µm in diameter), which contain the majority of cfs-mRNA, were enriched by a microfilter. Protein complexes, which most cfs-miRNA is bound with, were enriched by an ultrafilter. Harvesting the complexes or microvesicles, in which RNAs exist, avoided the influence of other components in human semen, thus favoring RNA isolation and purification. This new method can efficiently isolate cfs-mRNA and cfs-miRNA separately based on their physical nature, with high RNA purity, and low DNA contamination. It may also be applied or modified to isolate cell-free RNAs in other fluids.

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Paternal contributors in recurrent pregnancy loss: Cues from comparative proteome profiling of seminal extracellular vesicles

Jena

Nayak

Kumar

et al. 2020

Molecular Reproduction Devel

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Recent evidence entail paternal factors as plausible contributors in spontaneous recurrent pregnancy loss (RPL). Seminal extracellular vesicles secreted from cells of male reproductive tract carry regulatory proteins and RNAs. They are proposed to regulate sperm maturation and function while their fusion to endometrial stromal cells helps in decidualization. Nevertheless, the mechanism(s) involved in these processes are poorly understood. This study aims at elucidating the molecular basis of paternal contribution by comparative proteomics (label-free LC-MS/MS) of isolated seminal extracellular vesicles from fertile men and partners of patients with RPL (n = 21 per group). Bioinformatics analysis revealed the identified differentially expressed proteins to be involved in DNA replication, recombination and repair, gene expression, cellular assembly and organization, cell death, and survival. Major disease pathways affected were identified as developmental, hereditary, and immunological disorders. Of the three identified hub genes regulating the above disease pathways, two (HNRNPC and HNRNPU) are overexpressed while RUVBL1 is underexpressed along with over expression of HIST1H1C, DDX1, surmising defective chromatin packaging, and histone removal in spermatozoa resulting in improper expression in paternal genes thereby leading to abnormal embryo development. Besides, alteration in GSTP1 expression points oxidative predominance in RPL group. Differential expression of C3, C4a/C4b, CFB, and GDF 15 may be involved in altered maternal immune response to paternal antigens resulting in impaired decidualization.

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Quick recovery and characterization of cell-free DNA in seminal plasma of normozoospermia and azoospermia: implications for non-invasive genetic utilities

Cited by 52 publications

References 18 publications

Isolation of Cell-Free DNA from Seminal Fluid

Isolation of Cell-Free DNA from Seminal Fluid

Recovery of cell‐free mRNA and microRNA from human semen based on their physical nature

Paternal contributors in recurrent pregnancy loss: Cues from comparative proteome profiling of seminal extracellular vesicles

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