Use of fetal bovine serum to protect against zona hardening during preparation of mouse oocytes for cryopreservation

George, Martin A.; Johnson, Martin H.; Vincent, Chan

doi:10.1093/oxfordjournals.humrep.a137659

Cited by 23 publications

(12 citation statements)

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“…By contrast, if the medium was supplemented with FCS, FCS did not prevent premature CG exocytosis but did inhibit ZP conversion [21]. Also, in mice, DMSO can induce CG exocytosis and zona hardening, and prevent sperm penetration of oocytes [57,58]. However, when FCS was included in the medium, it also prevented DMSO-induced ZP hardening and ZP2 conversion without preventing CG exocytosis [57].…”

Section: Discussionmentioning

confidence: 96%

Parthenogenetic Activation of Pig Oocytes with Calcium Ionophore and the Block to Sperm Penetration after Activation1

Wang

Macháty

Abeydeera

et al. 1998

Biology of Reproduction

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Calcium ionophore A23187 can parthenogenetically activate oocytes in many animals. The present study was designed to analyze functionally the mechanism of A23187 activation of pig oocytes matured in vitro. In experiment 1, effects of the concentration of A23187 on intracellular calcium transients, cortical granule (CG) exocytosis, nuclear activation, and zona reaction, which was determined by zona hardening and sperm penetrability, were examined. Cumulus-free oocytes were exposed to 0-100 microM A23187 for 5 min. It was found that the amplitude of the intracellular calcium transients, percentage of CG exocytosis, and percentage of pronuclear formation were increased in a concentration-dependent manner. The time for dissolution of zona pellucida (ZP) was increased in the oocytes treated with 25-100 microM A23187. Penetration of ZP-intact oocytes by spermatozoa was decreased and only 3-4% of oocytes were penetrated by spermatozoa after 50-100 microM A23187 treatment. In experiment 2, oocytes were treated with 100 microM A23187 for 5 min and then cultured for 10 min or 3.5 h before insemination. No difference in penetration rates was observed between the two groups of oocytes (12.0% vs. 12.2%), but the penetration rates were significantly lower than those in controls (85.2% vs. 82.4%). In experiment 3, treatment of oocytes with 100 microM A23187 for 5 min was followed by removal of the ZP from a portion of the oocytes. ZP-intact and ZP-free oocytes were then inseminated for examination of sperm penetration. One of 65 (2%) oocytes with ZP and 48 of 52 (92%) oocytes without ZP were penetrated by spermatozoa. These results indicate that activation of pig oocytes by A23187 is the result of A23187-induced intracellular calcium increase and that A23187-induced cortical reaction can prevent sperm penetration of ZP-intact oocytes, but not ZP-free oocytes.

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Section: Discussionmentioning

confidence: 96%

Parthenogenetic Activation of Pig Oocytes with Calcium Ionophore and the Block to Sperm Penetration after Activation1

Wang

Macháty

Abeydeera

et al. 1998

Biology of Reproduction

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“…Previous work has shown that serum is a useful additive for increasing the cryopreservation efficiency of various cells and tissues [45, 46]. However, serum is not suitable for human clinical applications because of its lack of definition and potential to carry harmful agents such as viruses [12, 47].…”

Section: Discussionmentioning

confidence: 99%

Effect of Antioxidants and Apoptosis Inhibitors on Cryopreservation of Murine Germ Cells Enriched for Spermatogonial Stem Cells

Kim

Lee

et al. 2016

PLoS ONE

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Spermatogonial stem cells (SSCs) are germline stem cells that serve as the foundation of spermatogenesis to maintain fertility throughout a male’s lifetime. To treat male infertility using stem cell banking systems and transplantation, it is important to be able to preserve SSCs for long periods of time. Therefore, this study was conducted to develop an optimal cryopreservation protocol for SSCs using antioxidants and apoptosis inhibitors in freezing medium. No differences were observed compared to controls when SSCs were cryopreserved in the presence of apoptosis inhibitors by themselves. However, mouse germ cells cryopreserved in basal medium containing the antioxidant hypotaurine (14 mM) resulted in significantly greater proliferation potential and mitochondrial activity. Furthermore, treatment groups with combinations containing 200 mM trehalose and 14 mM hypotaurine showed higher proliferation rates compared to controls. In addition, several serum free conditions were evaluated for SSC cryopreservation. Treatment media containing 10% or 20% knockout serum replacement resulted in similar cryopreservation results compared to media containing FBS. SSC transplantation was also performed to confirm the functionality of SSCs frozen in 14 mM hypotaurine. Donor SSCs formed normal spermatogenic colonies and sperm in the recipient testis. These data indicate that inclusion of 14 mM hypotaurine in cryopreservation media is an effective way to efficiently cryopreserve germ cells enriched for SSCs and that knockout serum replacement can replace FBS in germ cell cryopreservation media.

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“…It also is questionable whether most follicular oocytes are capable of completing cytoplasmic maturation in vitro [16], a crucial prerequisite in other mammals for oocyte activation, fertilization, and development to the blastocyst stage [26]. Additionally, reduced IVM/IVF efficiency may be related to premature zona hardening, a response to fertilization triggered by cortical granule content release to prevent polyspermy [27], which may be brought on prematurely by in vitro culture [28,29]. Although zona hardening has never been confirmed in the cat, mechanically piercing the cat zona increases penetration in vitro by conspecific sperm [30].…”

Section: Introductionmentioning

confidence: 99%

Circannual Variations in Intraovarian Oocyte but Not Epididymal Sperm Quality in the Domestic Cat1

Spindler

Wildt

1999

Biology of Reproduction

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Ovaries and testes were collected throughout the year from domestic cats spayed and neutered at local veterinary clinics. Fresh oocytes recovered from minced ovaries were subjected to in vitro maturation and then stained to determine stage of maturation or were inseminated with conspecific sperm. The cauda and corpus regions of each epididymis were dissected into pieces and placed in medium; 30 min later, the epididymal tissue was removed, the medium centrifuged, and the sperm pellet resuspended. Samples were assessed for total sperm count and sperm motility traits, morphology, acrosomal integrity, and ability to penetrate cat oocytes in vitro. Fewer excellent (grade I) oocytes were recovered per ovarian pair during September-November (mean +/- SEM, 19.2 +/- 2.1%) than during January-July (36.8 +/- 3.6%, p < 0.05), while the remaining months had intermediate percentages of grade I oocytes (p > 0.05). A high percentage of oocytes recovered from November-April completed nuclear maturation (64.3 +/- 6.8%), which was different (p < 0.05) from the values for May-July (32.2 +/- 3.8%) and August-October (10.4 +/- 2.9%). Percentage of oocytes with bound sperm was lowest (p < 0.01) in September and October (32.0 +/- 3.1%) compared to February and March (91.4 +/- 1.7%). Percentage of oocytes with sperm within the perivitelline space was highest (p < 0.05) in May-August (33.8 +/- 4.6%) compared to all other months. In contrast, the period of highest (p < 0.01) fertilization (i.e., >/= 4-cell embryo formation) was March-April (51.7 +/- 3.1%) as compared to May-July (17.2 +/- 1.8%) or November-January (12.4 +/- 2.6%). Negligible numbers of oocytes recovered during August-October developed beyond the 2-cell stage (1.1 +/- 0.3%). Blastocyst development from cleaved embryos was highest during February-April (44.3 +/- 2.3%) and lowest during August-October (0.6 +/- 0.1%; p < 0.01). Sperm recovered from the epididymides throughout the year did not differ (p > 0.05) in concentration or in any of the motility, structural, or functional variables evaluated. In summary, cat oocyte nuclear maturation in vitro is depressed during August-October, and the ability to form cleaved embryos remains low even when the capacity to achieve nuclear maturation is relatively high (November-January and May-July). In contrast, male cats are capable of consistently producing viable, progressively motile sperm throughout the year.

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Use of fetal bovine serum to protect against zona hardening during preparation of mouse oocytes for cryopreservation

Cited by 23 publications

References 0 publications

Parthenogenetic Activation of Pig Oocytes with Calcium Ionophore and the Block to Sperm Penetration after Activation1

Parthenogenetic Activation of Pig Oocytes with Calcium Ionophore and the Block to Sperm Penetration after Activation1

Effect of Antioxidants and Apoptosis Inhibitors on Cryopreservation of Murine Germ Cells Enriched for Spermatogonial Stem Cells

Circannual Variations in Intraovarian Oocyte but Not Epididymal Sperm Quality in the Domestic Cat1

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