Cryopreservation-thawing of fractionated human spermatozoa and plasma membrane translocation of phosphatidylserine

Duru, Namık Kemal; Morshedi, Mahmood; Schuffner, Alessandro; Oehninger, Sergio

doi:10.1016/s0015-0282(00)01694-0

Cited by 87 publications

(68 citation statements)

References 24 publications

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“…A substantial fraction of AnVϩ/PIϪ BM CD34ϩ cells after different cell handling procedures has certainly not passed the point-ofno-return in the process of PCD, as the cells retained proliferative capacity, although to a lower extent. In analogy, cryopreservation and thawing of human spermatozoa (42) were associated with the induction of membrane PS translocation and high post-thaw levels of AnV were found even in the fractions with high sperm motility.…”

Section: Discussionmentioning

confidence: 92%

The dynamic process of apoptosis analyzed by flow cytometry using Annexin‐V/propidium iodide and a modified in situ end labeling technique

Span¹,

Pennings

Vierwinden

et al. 2001

Cytometry

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BackgroundTo study the apoptotic process in time, we used the following flow cytometric (FCM) techniques: phosphatidylserine (PS) translocation by Annexin‐V (AnV), DNA fragmentation by in situ end labeling (ISEL), and propidium iodide (PI) staining. Because PS translocation is assumed to be an early feature of programmed cell death (PCD), we questioned if AnV positivity implies inevitable cell death.MethodsApoptosis was induced in Jurkat cells by γ‐irradiation, incubation with camptothecin (CPT), or cytosine β‐D‐arabinofuranoside (Ara‐C). At different time intervals, PCD was quantified by AnV/PI and ISEL. To analyze the influence of cell handling procedures on PCD, we applied these three FCM techniques on CD34+ bone marrow (BM) stem cells after selection and after a freeze‐thaw procedure. Various AnV/PI− CD34+ fractions were cultured in a single‐cell single‐well (SCSW) assay.ResultsJurkat cells under three different detrimental conditions showed essentially the same pattern of apoptosis in time. Initially developed AnV+/PI− cells subsequently (within 1 h) showed ISEL positivity, after which they turned into AnV+/PI++ cells with even higher levels of ISEL positivity (80–90%). Eventually, they lost some of their PI and ISEL positivity and formed the AnV+/PI+ fraction. Cell handling of CD34+ cells caused high and variable AnV+/PI− fractions (overall range 23–62%). Within total AnV+ and AnV+/PI− populations, only a minority of CD34+ cells showed ISEL positivity (range 4–8% and 0.8–6%, respectively). Different fractions of AnV+/PI− CD34+ cells did have clonogenic capacity.ConclusionsPCD of cell suspensions in vitro can be followed accurately in time by these three FCM techniques. PS translocation is followed rapidly (within 1 h) by oligo‐nucleosomal DNA fragmentation, after which cell (and nuclear) membrane leakage occurs. Detection of PS asymmetry by AnV‐fluorescein isothiocyanate (FITC) is not always associated with (inevitable) apoptosis, as can be concluded from the proliferative capacity of AnV+ /PI− CD34+ cells in the SCSW assay. Cytometry 47:24–31, 2002. © 2001 Wiley‐Liss, Inc.

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Section: Discussionmentioning

confidence: 92%

The dynamic process of apoptosis analyzed by flow cytometry using Annexin‐V/propidium iodide and a modified in situ end labeling technique

Span¹,

Pennings

Vierwinden

et al. 2001

Cytometry

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“…Recent studies demonstrated that a variable proportion of live, ejaculated spermatozoa under in vitro incubation conditions shows PS externalization [24,42]. Since we already knew that sorted dual-stained spermatozoa die rapidly after incubation at 37-39…”

Section: Discussionmentioning

confidence: 99%

Sire Effect and Sperm Apoptosis on Bovine Embryonic Cleavage and Subsequent In Vitro Embryo Development

Chaveiro

Santos

Carvalhais

et al. 2010

SRX Veterinary Science

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The present study was conducted to evaluate sire effect on the kinetics of in vitro cleavage and further embryonic development after in vitro fertilization (IVF) using bulls of an Azorean rare breed called "Ramo Grande" (RG), and examine whether a relationship exists between bull's sperm apoptosis and sire in vitro fertility. Results showed that cleavage and blastocyst rates were statistically different between bulls (P < .05), being possible to notice different ability to produce embryos with good development competence (P < .05), as assessed by the proportion of fertilized oocytes that reached blastocyst stage. The proportion of blastocysts that continued the development to hatched blastocysts stage ranged from 9.1 to 87.5% (P < .05). Differences in sperm apoptosis increased during the swim-up procedure, which was statistically different (P < .05) between bulls, being negatively correlated with the ability of those bulls to fertilize oocytes and resulting embryos to develop to hatched blastocysts (R = −0.96, P ≤ .01).In conclusion, this study clearly demonstrated great differences between bulls, concerning apoptosis levels, which were negatively related to bull's ability to produce good quality embryos after IVF.

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“…Similarly, mechanical stress in other species as different as deer and dog does not lead to DNA damage [57,58], which suggests that mammalian sperm DNA can withstand some substantial amount of artificial physical stress and must also be very robust to any naturally occurring mechanical stress. By contrast, cryopreservation can lead to sperm DNA damage in some species such as ram, boar and alpaca [53,59,60], but not in others such as human, horse, bull and dog [58,[61][62][63].…”

Section: Discussionmentioning

confidence: 97%

A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation

et al. 2016

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Sperm competition, a prevalent evolutionary process in which the spermatozoa of two or more males compete for the fertilization of the same ovum, leads to morphological and physiological adaptations, including increases in energetic metabolism that may serve to propel sperm faster but that may have negative effects on DNA integrity. Sperm DNA damage is associated with reduced rates of fertilization, embryo and fetal loss, offspring mortality, and mutations leading to genetic disease. We tested whether high levels of sperm competition affect sperm DNA integrity. We evaluated sperm DNA integrity in 18 species of rodents that differ in their levels of sperm competition using the sperm chromatin structure assay. DNA integrity was assessed upon sperm collection, in response to incubation under capacitating or non-capacitating conditions, and after exposure to physical and chemical stressors. Sperm DNA was very resistant to physical and chemical stressors, whereas incubation in noncapacitating and capacitating conditions resulted in only a small increase in sperm DNA damage. Importantly, levels of sperm competition were positively associated with sperm DNA fragmentation across rodent species. This is the first evidence showing that high levels of sperm competition lead to an important cost in the form of increased sperm DNA damage.

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Cryopreservation-thawing of fractionated human spermatozoa and plasma membrane translocation of phosphatidylserine

Cited by 87 publications

References 24 publications

The dynamic process of apoptosis analyzed by flow cytometry using Annexin‐V/propidium iodide and a modified in situ end labeling technique

The dynamic process of apoptosis analyzed by flow cytometry using Annexin‐V/propidium iodide and a modified in situ end labeling technique

Sire Effect and Sperm Apoptosis on Bovine Embryonic Cleavage and Subsequent In Vitro Embryo Development

A cost for high levels of sperm competition in rodents: increased sperm DNA fragmentation

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