Optimizing human semen cryopreservation by reducing test vial volume and repetitive test vial sampling

TY - JOUR

T1 - Optimizing human semen cryopreservation by reducing test vial volume and repetitive test vial sampling

AU - Jensen, Christian F S

AU - Ohl, Dana A

AU - Parker, Walter R

AU - da Rocha, Andre M

AU - Keller, Laura M

AU - Schuster, Timothy G

AU - Sonksen, Jens

AU - Smith, Gary D

N1 - Copyright © 2015. Published by Elsevier Inc.

PY - 2015/3/1

Y1 - 2015/3/1

N2 - Objective To investigate optimal test vial (TV) volume, utility and reliability of TVs, intermediate temperature exposure (-88°C to -93°C) before cryostorage, cryostorage in nitrogen vapor (VN2) and liquid nitrogen (LN2), and long-term stability of VN2 cryostorage of human semen. Design Prospective clinical laboratory study. Setting University assisted reproductive technology (ART) laboratory. Patient(s) A total of 594 patients undergoing semen analysis and cryopreservation. Intervention(s) Semen analysis, cryopreservation with different intermediate steps and in different volumes (50-1,000 μL), and long-term storage in LN2 or VN2. Main Outcome Measure(s) Optimal TV volume, prediction of cryosurvival (CS) in ART procedure vials (ARTVs) with pre-freeze semen parameters and TV CS, post-thaw motility after two- or three-step semen cryopreservation and cryostorage in VN2 and LN2. Result(s) Test vial volume of 50 μL yielded lower CS than other volumes tested. Cryosurvival of 100 μL was similar to that of larger volumes tested. An intermediate temperature exposure (-88°C to -93°C for 20 minutes) during cryopreservation did not affect post-thaw motility. Cryosurvival of TVs and ARTVs from the same ejaculate were similar. Cryosurvival of the first TV in a series of cryopreserved ejaculates was similar to and correlated with that of TVs from different ejaculates within the same patient. Cryosurvival of the first TV was correlated with subsequent ARTVs. Long-term cryostorage in VN2 did not affect CS. Conclusion(s) This study provides experimental evidence for use of a single 100 μL TV per patient to predict CS when freezing multiple ejaculates over a short period of time (<10 days). Additionally, semen cryostorage in VN2 provides a stable and safe environment over time.

AB - Objective To investigate optimal test vial (TV) volume, utility and reliability of TVs, intermediate temperature exposure (-88°C to -93°C) before cryostorage, cryostorage in nitrogen vapor (VN2) and liquid nitrogen (LN2), and long-term stability of VN2 cryostorage of human semen. Design Prospective clinical laboratory study. Setting University assisted reproductive technology (ART) laboratory. Patient(s) A total of 594 patients undergoing semen analysis and cryopreservation. Intervention(s) Semen analysis, cryopreservation with different intermediate steps and in different volumes (50-1,000 μL), and long-term storage in LN2 or VN2. Main Outcome Measure(s) Optimal TV volume, prediction of cryosurvival (CS) in ART procedure vials (ARTVs) with pre-freeze semen parameters and TV CS, post-thaw motility after two- or three-step semen cryopreservation and cryostorage in VN2 and LN2. Result(s) Test vial volume of 50 μL yielded lower CS than other volumes tested. Cryosurvival of 100 μL was similar to that of larger volumes tested. An intermediate temperature exposure (-88°C to -93°C for 20 minutes) during cryopreservation did not affect post-thaw motility. Cryosurvival of TVs and ARTVs from the same ejaculate were similar. Cryosurvival of the first TV in a series of cryopreserved ejaculates was similar to and correlated with that of TVs from different ejaculates within the same patient. Cryosurvival of the first TV was correlated with subsequent ARTVs. Long-term cryostorage in VN2 did not affect CS. Conclusion(s) This study provides experimental evidence for use of a single 100 μL TV per patient to predict CS when freezing multiple ejaculates over a short period of time (<10 days). Additionally, semen cryostorage in VN2 provides a stable and safe environment over time.

KW - Calibration

KW - Cell Survival

KW - Cryopreservation

KW - Humans

KW - Male

KW - Semen

KW - Semen Analysis

KW - Semen Preservation

KW - Specimen Handling

KW - Sperm Retrieval

KW - Temperature

KW - Time Factors

U2 - 10.1016/j.fertnstert.2014.12.107

DO - 10.1016/j.fertnstert.2014.12.107

M3 - Journal article

C2 - 25585506

SN - 0015-0282

VL - 103

SP - 640-6.e1

JO - Fertility and Sterility

JF - Fertility and Sterility

IS - 3

ER -