Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad

Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad

Cryopreservation is a technique commonly used to preserve cells for long time storage. It is widely used in livestock industry to store male gametes in liquid nitrogen. In this study, three Bos taurus adult were used. Semen collected from these bulls were grouped into two; 1) treatment group whic...

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Main Author: Syed Mohamad, Syarifah Faezah
Format: Thesis
Language:English
Published: 2013
Subjects:
Magnetism
Magnetic fields
Online Access:http://ir.uitm.edu.my/id/eprint/15504/
http://ir.uitm.edu.my/id/eprint/15504/1/TM_SYARIFAH%20FAEZAH%20SYED%20MOHAMAD%20AS%2013_5.PDF
id uitm-15504
recordtype eprints
spelling uitm-155042016-11-18T03:56:35Z http://ir.uitm.edu.my/id/eprint/15504/ Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad Syed Mohamad, Syarifah Faezah Magnetism Magnetic fields Cryopreservation is a technique commonly used to preserve cells for long time storage. It is widely used in livestock industry to store male gametes in liquid nitrogen. In this study, three Bos taurus adult were used. Semen collected from these bulls were grouped into two; 1) treatment group which was subjected to Magnetic Activated Cell Sorting System (MACS) then cryopreserved, and 2) the untreated group which was directly cryopreserved without MACS. Prior to cryopreservation, sperm were incubated in Annexin V then placed in the column attached to the magnetic stand. Semen analysis was done by using the Computer Assisted Sperm Analysis software (CASA). Parameters included were sperm velocity (VAP, VCL, VSL; nm/s) and sperm progression (WOB, LIN, STR; %). Sperm viability was determined by using eosin-nigrosin staining technique. Effects of different thawing temperatures and times were analysed by using iQ™SYBR® Green assay for qPCR with GAPDH as the reference gene. Cryo-capacitation like damage was then identified using chlorotetracycline/Hoechst staining assay under blue-violet illumination of fluorescent microscope (excitation: 400-440 nm, emission: 470 nm). All statistical data were analysed using SPSS version 14; p-value was set at 0.05 as significant level. Findings revealed that thawing at 37°C for 30 seconds for MACS processed sample was most suitable compared to other thawing procedures. At 37°C for 30 seconds, it produced significant data of VCL nm/s, 133.31 [im/s ± 4.38 (p = 0.004), RNA concentration of 83.84 ng/ pi (p = 0.001), relative gene expression of HSP70, 0.097 ± 0.027 (p = 0.048) and CTC/Hoechst assay, 50.6% of F pattern (p = 0.045). Hence, we can speculate that the usage of MACS as the sperm preparation technique as able to enhance sperm cryosurvival rate after cryopreservation, however the fertilization potential of these sperm has yet to be determined. 2013 Thesis NonPeerReviewed text en http://ir.uitm.edu.my/id/eprint/15504/1/TM_SYARIFAH%20FAEZAH%20SYED%20MOHAMAD%20AS%2013_5.PDF Syed Mohamad, Syarifah Faezah (2013) Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad. Masters thesis, Universiti Teknologi MARA.
repository_type Digital Repository
institution_category Local University
institution Universiti Teknologi MARA
building UiTM Institutional Repository
collection Online Access
language English
topic Magnetism
Magnetic fields
spellingShingle Magnetism
Magnetic fields
Syed Mohamad, Syarifah Faezah
Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad
description Cryopreservation is a technique commonly used to preserve cells for long time storage. It is widely used in livestock industry to store male gametes in liquid nitrogen. In this study, three Bos taurus adult were used. Semen collected from these bulls were grouped into two; 1) treatment group which was subjected to Magnetic Activated Cell Sorting System (MACS) then cryopreserved, and 2) the untreated group which was directly cryopreserved without MACS. Prior to cryopreservation, sperm were incubated in Annexin V then placed in the column attached to the magnetic stand. Semen analysis was done by using the Computer Assisted Sperm Analysis software (CASA). Parameters included were sperm velocity (VAP, VCL, VSL; nm/s) and sperm progression (WOB, LIN, STR; %). Sperm viability was determined by using eosin-nigrosin staining technique. Effects of different thawing temperatures and times were analysed by using iQ™SYBR® Green assay for qPCR with GAPDH as the reference gene. Cryo-capacitation like damage was then identified using chlorotetracycline/Hoechst staining assay under blue-violet illumination of fluorescent microscope (excitation: 400-440 nm, emission: 470 nm). All statistical data were analysed using SPSS version 14; p-value was set at 0.05 as significant level. Findings revealed that thawing at 37°C for 30 seconds for MACS processed sample was most suitable compared to other thawing procedures. At 37°C for 30 seconds, it produced significant data of VCL nm/s, 133.31 [im/s ± 4.38 (p = 0.004), RNA concentration of 83.84 ng/ pi (p = 0.001), relative gene expression of HSP70, 0.097 ± 0.027 (p = 0.048) and CTC/Hoechst assay, 50.6% of F pattern (p = 0.045). Hence, we can speculate that the usage of MACS as the sperm preparation technique as able to enhance sperm cryosurvival rate after cryopreservation, however the fertilization potential of these sperm has yet to be determined.
format Thesis
author Syed Mohamad, Syarifah Faezah
author_facet Syed Mohamad, Syarifah Faezah
author_sort Syed Mohamad, Syarifah Faezah
title Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad
title_short Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad
title_full Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad
title_fullStr Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad
title_full_unstemmed Study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and HSP70 gene expression / Syarifah Faezah Syed Mohamad
title_sort study on optimization of cryopreserved bovine spermatozoa using magnetic activated cell sorting system on viability, motility and hsp70 gene expression / syarifah faezah syed mohamad
publishDate 2013
url http://ir.uitm.edu.my/id/eprint/15504/
http://ir.uitm.edu.my/id/eprint/15504/1/TM_SYARIFAH%20FAEZAH%20SYED%20MOHAMAD%20AS%2013_5.PDF
first_indexed 2023-09-18T22:54:00Z
last_indexed 2023-09-18T22:54:00Z
_version_ 1777417731889430528

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