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Comparison of the effect of different media on the clinical outcomes of the density-gradient centrifugation/swim-up and swim-up methods.

Kim EK, Kim EH, Kim EA, Lee KA, Shin JE, Kwon H - Clin Exp Reprod Med (2015)

Bottom Line: Poor responders with less than three mature oocytes were excluded.There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes.We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

View Article: PubMed Central - PubMed

Affiliation: Fertility Center, CHA Bundang Medical Center, Seongnam, Korea.

ABSTRACT

Objective: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF.

Methods: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123).

Results: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates.

Conclusion: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

No MeSH data available.


Related in: MedlinePlus

Schematic diagram for preparing sperm with the density gradient centrifugation-swim-up (DGC-SUP) and swim-up (SUP) methods using Ham's F-10 medium and sperm washing medium (SWM). WBC, white blood cell; HPF, high power field; SSS, synthetic serum substitute.
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Figure 1: Schematic diagram for preparing sperm with the density gradient centrifugation-swim-up (DGC-SUP) and swim-up (SUP) methods using Ham's F-10 medium and sperm washing medium (SWM). WBC, white blood cell; HPF, high power field; SSS, synthetic serum substitute.

Mentions: Discontinuous DGC was performed using a sterile pipette. The lower layer (90% Isolate, Irvine Scientific, Santa Ana, CA, USA) was first transferred into a conical centrifuge tube. Using a new sterile pipette, the upper layer (50% Isolate, Irvine Scientific) was gently dispensed on top of the lower layer. A liquefied 2.0 mL semen sample was then placed on top of the upper layer and the tube was centrifuged for 10 minutes at 300 g. This process was repeated using additional tubes until the entire sample of ejaculate was processed. The upper and lower layers were carefully aspirated without disturbing the pellet and discarded. Using a transfer pipette, 4.0 mL of Ham's F-10 medium (Irvine Scientific) with 0.5% synthetic serum substitute (Irvine Scientific) or 4.0 mL of HEPES-buffered SWM (Irvine Scientific) was added onto the pellet and the resuspended pellet was centrifuged for five minutes at 250 g. The washing procedure was repeated twice. The supernatant was then removed and the pellet was suspended in 0.5 mL of Ham's F-10 medium or SWM, which was gently layered on top of the pellet, and the tube was inclined at an angle of 45 degrees and incubated at 35℃ for at least 30-40 minutes. After the incubation period, a sterile pipette was used to aspirate the supernatant and transfer it to a sterile 5-mL conical tube. Sperm count and motility were determined in the recovered fractions (Figure 1).


Comparison of the effect of different media on the clinical outcomes of the density-gradient centrifugation/swim-up and swim-up methods.

Kim EK, Kim EH, Kim EA, Lee KA, Shin JE, Kwon H - Clin Exp Reprod Med (2015)

Schematic diagram for preparing sperm with the density gradient centrifugation-swim-up (DGC-SUP) and swim-up (SUP) methods using Ham's F-10 medium and sperm washing medium (SWM). WBC, white blood cell; HPF, high power field; SSS, synthetic serum substitute.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390677&req=5

Figure 1: Schematic diagram for preparing sperm with the density gradient centrifugation-swim-up (DGC-SUP) and swim-up (SUP) methods using Ham's F-10 medium and sperm washing medium (SWM). WBC, white blood cell; HPF, high power field; SSS, synthetic serum substitute.
Mentions: Discontinuous DGC was performed using a sterile pipette. The lower layer (90% Isolate, Irvine Scientific, Santa Ana, CA, USA) was first transferred into a conical centrifuge tube. Using a new sterile pipette, the upper layer (50% Isolate, Irvine Scientific) was gently dispensed on top of the lower layer. A liquefied 2.0 mL semen sample was then placed on top of the upper layer and the tube was centrifuged for 10 minutes at 300 g. This process was repeated using additional tubes until the entire sample of ejaculate was processed. The upper and lower layers were carefully aspirated without disturbing the pellet and discarded. Using a transfer pipette, 4.0 mL of Ham's F-10 medium (Irvine Scientific) with 0.5% synthetic serum substitute (Irvine Scientific) or 4.0 mL of HEPES-buffered SWM (Irvine Scientific) was added onto the pellet and the resuspended pellet was centrifuged for five minutes at 250 g. The washing procedure was repeated twice. The supernatant was then removed and the pellet was suspended in 0.5 mL of Ham's F-10 medium or SWM, which was gently layered on top of the pellet, and the tube was inclined at an angle of 45 degrees and incubated at 35℃ for at least 30-40 minutes. After the incubation period, a sterile pipette was used to aspirate the supernatant and transfer it to a sterile 5-mL conical tube. Sperm count and motility were determined in the recovered fractions (Figure 1).

Bottom Line: Poor responders with less than three mature oocytes were excluded.There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes.We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

View Article: PubMed Central - PubMed

Affiliation: Fertility Center, CHA Bundang Medical Center, Seongnam, Korea.

ABSTRACT

Objective: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF.

Methods: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123).

Results: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates.

Conclusion: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

No MeSH data available.


Related in: MedlinePlus