Is fine morphology of the human sperm nuclei affected by in vitro incubation at 37°C?

Pe’er, Sara; Eltes, F.; Berkovitz, Arie; Yehuda, Ronen; Itsykson, Pavel; Bartoov, B.

doi:10.1016/j.fertnstert.2007.01.069

Cited by 81 publications

(59 citation statements)

References 23 publications

(26 reference statements)

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“…In addition, Hammadeh et al [27] reported a significant increase in spermatozoa with decondensed chromatin upon in vitro incubation at 37ºC for 24 h. Furthermore, it has been suggested that prolonged sperm cell manipulations should be performed at RT (21ºC) rather than at 37ºC, because extended handling of spermatozoa at 37ºC is detrimental to fine sperm nuclear morphology [28]. In the present study, sperm motility decreased and %DFI increased in a time-dependent manner at both RT (25ºC) and 37ºC.…”

Section: Discussionmentioning

confidence: 99%

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Matsuura¹,

Takeuchi²,

Yoshida³

2010

Asian J Androl

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Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay (SCSA) parameters, that is, DNA fragmentation index (%DFI) and high DNA stainability (%HDS), were assessed on the fresh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation (DGC) was followed by swimup treatment in comparison with DGC alone (P < 0.01). Although the %DFI increased in a time-dependent manner with incubation both at room temperature (RT) and at 37ºC in air, the %DFI after 24 h at RT was significantly lower than that at 37ºC (P < 0.05). Incubation with 5% CO2 was effective in maintaining sperm motility (P < 0.01); however, it induced further elevation of %DFI (P < 0.001). Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.

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Section: Discussionmentioning

confidence: 99%

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Matsuura¹,

Takeuchi²,

Yoshida³

2010

Asian J Androl

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“…Les anomalies chromosomiques de nombre des spermatozdides, ou aneupld/dies, sont plus fr~quentes en cas de t~ratospermie majeure et monomorphe. Ces anomalies chromosomiques peuvent retentir sur le d6veloppement embryonnaire et favoriser la survenue de fausses couches [34,40].…”

Section: Impact De La Morphologie Des Spermatozo[des Sur La Fertilitrunclassified

Sperm morphology examination

Saïdi

Gruel

Roset-Blessman

et al. 2008

Androl.

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Mots cl6s : infertitit6, morphologie, spermatozoi'de, t6ratozoospermie I. INTRODUCTION L'exarnen du sperme (sperrnogramme -spermocytogramme) constitue une des ~tapes clefs dans rexploration du couple infertile. /'~jaculat normal contient des spermatozoi'des pr~sentant des variations importantes de la taille et de la forme de la t~te, de I'acrosome, de la pi(bce interm~diaire et du flagelle. Cette h6terog~n6it~ s'explique par le fait que le sperrnatozd[de, ceUule hautement diff~renci~e, est le r~sultat ultime d'un processus complexe : la spermatogen~se. Ce processus peut ~tre la cible de facteurs toxiques endog~nes ou exog~nes qui peuvent induire la production excessive de spermatozoides morphologiquement anormaux responsable d'une t~ratozoospermie. Une morphologie anormale des spermatozofdes peut comprornettre le pouvoir f6condant d'un sperme. Cependant, le profil morphologique du sperrne semble ~tre le param~tre sperrnatique le plus constant chez un homme [12,19,30,36].La morphologie du sperme a et~ reconnue comme le meilleur facteur pr~dictif de la fertilit~ naturelle ou de I'issue en inseminations intra ut~rines (IIU) ou en f6condation in vitro (FIV) classique [8,38]. Cependant, la majoritd des auteurs n'ont pas retrouv6 de relation entre la morphologie du sperrne et le succ~s de I'ICSI [20,36,31]. Quatre explications ont ~t~ propos~es. Premi~rement, une morphologie normale du spermatozoi'de est requise pour le passage des barri~res ovocytaires qui sont court-circuit~es par I'ICSI. Deuxi~mement, les spermatozdfdes portant des anomalies morphologiques ~videntes ne sont pas retenus pour rinjection. Troisi~mement, le spermocytogramme r~alis~ sur des ceUules randomisees de I'~jaculat n'est pas le reflet exact de la qualit6 du spermatozoi"de inject6. Quatri~mement, les Correspondance :Pr Nathalie RIVES -Laboratoire de Biologie de la I rue de Germont, CHU H6pitaux de Rouen, 76031, 158 anomalies ultrastructurales qui sont les seules associees au devenir de I'ICSI ne peuvent pas (~tre d~tect~es ni au grossissement xl00, ni au grossissement x200-400 [4,6]. L'analyse morphologique du sperme ou spermocytogramme est une dtude descriptive de la forme des spermatozofdes partir d'un ~chantillon repr6sentatif d'un 6jaculat. La morphologie n'est qu'un des param~tres qualitatifs de la spermatogen~se. Cette analyse est d~pendante des techniques d'imagerie utilisdes et des criteres d'interprdtation adopt6s pour ddfinir un spermatozo'ide morphologiquement normal. II. MICROSCOPIE OPTIQUEC'est la mdthode la plus utilisde et elle fait partie des examens de routine du fait de son faible coot et de sa simplicitY. L'analyse morphologique des spermatozoi'des est faite partir de preparations fixdes et colordes examinees en lumi~re transmise au grossissement final xl000 (objectif 100 immersion). Mdthodes de colorationPlusieurs colorations sont possibles : Giemsa, coloration de Papanicolaou modifiee pour les spermatozoides (PAP), coloration de Bruyan-Leishman ou encore coloration de SchorT. D'apr~s certains auteurs, la coloration de...

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“…The spermatozoa were assessed at room temperature, based on MSOME criteria, using an inverted microscope (Leica DMI6000B, Wetzlar, Germany) equipped with high power differential interference contrast optics to achieve a final electronic magnification of 36650, as described by Peer et al 17 At least 20 spermatozoa, classified as normal according to MSOME criteria (smooth, symmetric and oval configuration of the sperm head with an average length of 4.7560.28 mm and average width of 3.2860.20 mm, normal acrosome and postacrosomal lamina with normal neck and tail, and no extrusion or invagination of the nuclear chromatin mass), with small nuclear vacuoles occupying less than 4% of the nuclear area, 9,18 were collected in a small drop of fertilization medium. In the cases where the best spermatozoa could not be found, the second-best spermatozoa (those with a single specific nuclear malformation and without a large nuclear vacuole) were chosen as described by Berkovitz et al 19 In the PICSI group, a Petri dish coated with HA (Biocoat, Fort Washington, PA, USA) was used for sperm selection, as described by Huszar et al 12 In brief, a drop of sperm suspension was placed close to the edge of the HA spot, and the spermatozoa were allowed to migrate to the drop of HA.…”

Section: Methods Of Sperm Selectionmentioning

confidence: 99%

No difference in high-magnification morphology and hyaluronic acid binding in the selection of euploid spermatozoa with intact DNA

Mongkolchaipak¹,

Vutyavanich²

2013

Asian J Androl

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In this study, we compared conventional sperm selection with high-magnification morphology based on the motile sperm organellar morphology examination (MSOME) criteria, and hyaluronic acid (HA) binding for sperm chromosome aneuploidy and DNA fragmentation rates. Semen from 50 severe male factor cases was processed through density gradient centrifugation, and subjected to sperm selection by using the conventional method (control), high magnification at 36650 or HA binding. Aneuploidy was detected by fluorescence in situ hybridization with probes for chromosomes 13, 18, 21, X and Y, and DNA fragmentation by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) method. Spermatozoa selected under high-magnification had a lower DNA fragmentation rate (2.6% vs. 1.7%; P50.032), with no significant difference in aneuploidy rate (0.8% vs 0.7%; P50.583), than those selected by the HA binding method. Spermatozoa selected by both methods had much lower aneuploidy and DNA fragmentation rate than the controls (7% aneuploidy and 26.8% DNA fragmentation rates, respectively). In the high-magnification group, the aneuploidy rate was lower when the best spermatozoa were selected than when only the second-best spermatozoa were available for selection, but the DNA fragmentation rate was not different. In conclusion, sperm selection under high magnification was more effective than under HA binding in selecting spermatozoa with low DNA fragmentation rate, but the small difference (0.9%) might not be clinically meaningful. Both methods were better than the conventional method of sperm selection. Keywords: aneuploidy; DNA fragmentation; hyaluronic acid binding; intracytoplasmic sperm injection (ICSI); intracytoplasmic morphologically selected sperm injection (IMSI); motile sperm organellar morphology examination; physiologic intracytoplasmic sperm injection INTRODUCTION Intracytoplasmic sperm injection (ICSI) is a very effective method for the treatment of severe male factor infertility. It allows the use of a single motile spermatozoon to fertilise an oocyte. 1 The technique is so effective that fertilization of an oocyte can be achieved even with the use of a spermatozoon with severe DNA fragmentation. 2 Men with severe male factor infertility are known to have a higher frequency of sperm aneuploidies and DNA fragmentation than fertile men. [3][4][5][6][7][8] Although the consequences of inseminating oocytes with abnormal spermatozoa are not known for certain, there is increasing evidence that such circumstances may cause poor fertilization, defective pre-implantation embryonic development, and high rates of miscarriage and morbidity in the offspring, including childhood cancer. 8 One major challenge ICSI facing is, therefore, the selection of the best spermatozoon for micro-insemination.Many studies have shown that intracytoplasmic morphologicallyselected sperm injection (IMSI) can significantly increase the fertilization and pregnancy rate of ICSI, while decreasing its abortion rate. [9][10][11] The disadvantage...

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Is fine morphology of the human sperm nuclei affected by in vitro incubation at 37°C?

Cited by 81 publications

References 23 publications

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Sperm morphology examination

No difference in high-magnification morphology and hyaluronic acid binding in the selection of euploid spermatozoa with intact DNA

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