Sperm Collection of Differential Quality Using Density Gradient Centrifugation

Ahammad, Muslah Uddin; Jarrell, Zachery; Benson, Aaron

doi:10.3791/58833

Cited by 4 publications

(6 citation statements)

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“…Semen samples were collected from an additional 24 randomly selected AYP roosters by dorsoabdominal massage, and 4 pooled samples were generated by combining 6 individual samples. Three milliliters of each pooled sample were diluted 1:2 in PBS and subjected to Percoll density gradient centrifugation ( PDGC ) as described by Ahammad et al. (2018) in triplicate.…”

Section: Methodsmentioning

confidence: 99%

Characterization of sperm-associated antigen 6 expression in the reproductive tract of the domestic rooster (Gallus domesticus) and its impact on sperm mobility

et al. 2020

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Sperm mobility is a major determinant of sperm quality in the domesticated chicken ( Gallus domesticus ) and is therefore an area of interest for improving fertility. Sperm-associated antigen 6 ( SPAG6 ) is an important flagellar protein implicated to be necessary for flagellar function but negatively associated with rooster fertility. This study was aimed to characterize the expression of SPAG6 and investigate its utility as a protein biomarker of sperm mobility. By western analysis, relative SPAG6 abundances were compared between the testicular, epididymal, and vasal tissues and in sequentially maturing sperm. Immunocytochemistry techniques were used to detect localization of SPAG6 in chicken sperm. Last, western analysis was used to compare relative SPAG6 abundances in sperm of differing mobility. SPAG6 was found in higher abundance in epididymal tissues and in highest abundance in vasal tissues, relative to that of the testis. SPAG6 was also found to sequentially increase in abundance in maturing sperm. SPAG6 localizes between the axonemal central pair of microtubules in the sperm flagella, but it is also found in lower concentration in the acrosomal region. SPAG6 was not a significant predictor of sperm mobility. SPAG6 abundance, alone, is not a strong predictor of sperm mobility. Its impact on rooster fertility is likely unrelated to its impact on sperm mobility.

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Section: Methodsmentioning

confidence: 99%

Characterization of sperm-associated antigen 6 expression in the reproductive tract of the domestic rooster (Gallus domesticus) and its impact on sperm mobility

et al. 2020

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“…To perform assisted reproduction techniques (ART), sperm must first be processed by selecting for morphologically normal spermatozoa with high motility in order to maximize the concentration of good quality sperm. The preparation also removes cell debris, including epithelial cells and leukocytes (Malvezzi et al, 2014;Ahammad et al, 2018). The semen preparation procedure is therefore an important factor that directly affects the outcome of infertility treatment.…”

Section: Introductionmentioning

confidence: 99%

“…Semen is placed on top of media layered at different dilutions in a disposable centrifuge tube and, after centrifugation, each sperm cell will be located at the level of the gradient that corresponds to its density. As a result, highly motile, morphologically normal, and viable sperm form a pellet at the bottom of the tube, leukocytes and cells debris will be trapped at the interface between the seminal plasma and the more dilute upper phase, and morphologically abnormal sperm with low motility will be at the interface between the layers (Ahammad et al, 2018). This method is considered simple, reproducible, and efficient for isolating high quality sperm for use in ART and has been widely used in cases of normospermic patients (Ahammad et al, 2018;Jayaraman et al, 2012;Yang et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Initially, Percoll was widely used, which is composed of silica particles coated with polyvinylpyrrolidone (PVP). However, due to the toxicity of PVP, its use in humans has been discontinued (Mousset-Siméon et al, 2004;Ahammad et al, 2018). Since then, iodixanol or colloidal solutions of silica particles coated with silane have been tested as potential alternatives (Mousset-Siméon et al, 2004;Lee & Jee, 2019).…”

Section: Introductionmentioning

confidence: 99%

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Comparative sperm recovery rate after density gradient centrifugation with two media for in vitro fertilization

Fernandes¹,

Silva²,

Panizzon³

et al. 2022

JBRA

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Objective: To compare the efficacy of two density gradient centrifugation media for retrieving spermatozoa from semen samples by evaluating the total motile sperm count (TMSC) and the percentage recovery.Methods: Twenty-two men with different sperm counts participated in the study. The samples were divided into two equal aliquots and processed using the commercial ISolate Sperm Separation Medium (Irvine Scientific, United States) and the GV Gradiente (IngáMed, Brazil). After separation, samples were counted and evaluated for motile sperm recovery.Results: The mean TMSC in the fresh sample was 19.65±21.08 million/mL. After the ISolate separation the TMSC was 6.71±7.29 million/mL, and for the GV Gradiente it was 6.27±6.82 million/mL. The percentage of motile spermatozoa recovered was 36.47%±21.61 for ISolate and 35.22%±21.24 for the GV Gradiente (p>0.05). The samples from 6 oligospermic patients (27%) were evaluated separately and the TMSC for ISolate was 4.83±2.92 million/mL, and for the GV Gradiente, it was 4.16±3.12 million/mL (p=0.54). When evaluating only normospermic patient samples, the TMSC for ISolate was 9.05±7.29 million/mL, and for the GV Gradiente, it was 8.47±6.79 million/mL (p=0.83).Conclusions: There was no statistical difference in retrieving motile sperm using the GV Gradiente and the ISolate Separation Medium.

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“…Las muestras seminales se obtuvieron de los donadores con abstinencia sexual de ff.Primera separación de espermatozoides La separación de los gametos móviles entre la población total de espermatozoides del eyaculado se realizó por la técnica de gradiente, la solución de gradiente discontinuo de densidad es diseñado para obtener muestras limpias rápidamente, con altas concentraciones de espermatozoides móviles progresivos, tanto de semen saludable como de baja calidad, el cual consta de una capa superior de densidad del 50% y una capa inferior del 90% 61 .…”

Section: Imaterials Y Métodosunclassified

Evaluación funcional de espermatozoides posterior a la separación por columnas magnéticas

Morales

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Sperm Collection of Differential Quality Using Density Gradient Centrifugation

Cited by 4 publications

References 0 publications

Characterization of sperm-associated antigen 6 expression in the reproductive tract of the domestic rooster (Gallus domesticus) and its impact on sperm mobility

Characterization of sperm-associated antigen 6 expression in the reproductive tract of the domestic rooster (Gallus domesticus) and its impact on sperm mobility

Comparative sperm recovery rate after density gradient centrifugation with two media for in vitro fertilization

Evaluación funcional de espermatozoides posterior a la separación por columnas magnéticas

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