scholarly journals The Effect of Density Gradient Centrifugation and Magnetic-Activated Cell Sorting with the Combination of Both on Bovine Spermatozoa Kinematics, Viability and Acrosome Status

2017 ◽  
Vol 9 (13) ◽  
pp. 73
Author(s):  
Khairul Osman ◽  
Noraina Mohd Bakri ◽  
Farah Hanan Fathihah Jaffar ◽  
Nurul Atikah Osman ◽  
Siti Fatimah Ibrahim
Keyword(s):  
Density Gradient ◽  
Cell Sorting ◽  
Assisted Reproductive Technologies ◽  
Density Gradient Centrifugation ◽  
Semen Analysis ◽  
Gradient Centrifugation ◽  
Reproductive Technologies ◽  
Lower Percentage ◽  
Computer Assisted ◽  
Magnetic Activated Cell Sorting

Conventional and novel spermatozoa separation techniques have their own strength and weaknesses. Both are widely used in assisted reproductive technologies (ARTs). In this study, combinations of two separation methods against single separation methods were evaluated. Freshly ejaculated semen was collected and divided into five groups; control, density gradient centrifugation (DGC), magnetic activated cell sorting (MACS), DGC-MACS and MACS-DGC; based on the sequence of separation. Evaluation of spermatozoa kinematics, viability and acrosome status for each group were done by computer assisted semen analysis (CASA), hypo-osmotic swelling test (HOST) and chlorotetracycline (CTC) staining respectively. Results show that DGC yield the fastest spermatozoa velocity and highest percentage of viable spermatozoa fraction compared to other groups. However, they also produced a high percentage of acrosome-reacted spermatozoa. Spermatozoa isolated through DGC-MACS shows the highest percentage of non-capacitated spermatozoa. However, spermatozoa isolated through DGC-MACS yields slower spermatozoa velocity population and lower percentage of viable sperm compared to spermatozoa isolated through DGC. Isolation of spermatozoa through MACS-DGC yields slower population of spermatozoa velocity with a lower percentage of non-capacitated spermatozoa compared to spermatozoa isolated through DGC-MACS. DGC separation and its combination with MACS had yield different spermatozoa quality. This distinct characteristic is likely suitable for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) procedure respectively.

scholarly journals Magnetic-activated cell sorting before density gradient centrifugation improves recovery of high-quality spermatozoa

Andrology ◽  
2017 ◽  
Vol 5 (4) ◽  
pp. 776-782 ◽  
Author(s):  
T. S. Berteli ◽  
M. G. Da Broi ◽  
W. P. Martins ◽  
R. A. Ferriani ◽  
P. A. Navarro
Keyword(s):  
Density Gradient ◽  
Cell Sorting ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
High Quality ◽  
Magnetic Activated Cell Sorting

scholarly journals Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing

2006 ◽  
Vol 48 (4) ◽  
pp. 201-206 ◽  
Author(s):  
Cynthia Liliane Motta do Canto ◽  
Aluisio C. Segurado ◽  
Cláudio Pannut ◽  
Agnaldo Cedenho ◽  
Miguel Srougi ◽  
...  
Keyword(s):  
Density Gradient ◽  
Assisted Reproductive Technologies ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Reproductive Technologies ◽  
Hcv Rna ◽  
Rna Detection ◽  
Hiv Rna ◽  
Hiv Serodiscordant Couples ◽  
Before And After

INTRODUCTION: Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests. METHODS: After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen. RESULTS: In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions. CONCLUSIONS: Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

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