Freeze-Dried Sperm Fertilization Leads to Full-Term Development in Rabbits1

Liu, Ji‐Long; Kusakabe, Hirokazu; Chang, Ching-Chien; Suzuki, Hiroyuki; Schmidt, D; Julian, Marina; Pfeffer, Robert; Bormann, Charles L.; Tian, X. Cindy; Yanagimachi, Ryuzo; Yang, Xiangzhong

doi:10.1095/biolreprod.103.025957

Cited by 164 publications

(142 citation statements)

References 31 publications

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“…The value was obviously higher than in the other groups. A possible reason is that the plasma membrane of the freeze-dried spermatozoa was completely destroyed [7,11,15], which could lead to a more rapid contact between the sperm nuclei and the ooplasm.…”

Section: Discussionmentioning

confidence: 99%

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Pronuclear formation of freeze-dried canine spermatozoa microinjected into mouse oocytes

Watanabe

Asano

Abe

et al. 2009

J Assist Reprod Genet

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Purpose The aim of the present study was to investigate the fertilizing capacity of fresh, frozen-thawed and freeze-dried canine spermatozoa. Methods After canine spermatozoa were injected into mouse oocytes, the rates of oocyte activation, male pronuclear formation and chromosomal aberrations were investigated. Results The rates of oocyte activation were comparable (90.6-100%), no matter the sperm type injected. The percentage of male pronuclear formation was higher (P< 0.001) in the freeze-dried spermatozoa (92.3%) than the fresh (61.5%) and frozen-thawed (69.2%) spermatozoa. However, the chromosomal damage in the oocytes injected with freezedried spermatozoa was higher (72.9%: P < 0.001) than with fresh (26.9%) and frozen-thawed (21.4%) spermatozoa. Conclusions These data indicate using mouse oocytes that freeze-dried canine spermatozoa may potentially fertilize canine oocytes although chromosomal damage is frequently generated.

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Section: Discussionmentioning

confidence: 99%

“…As one of the cryopreservation methods in mammalian spermatozoa, freeze-drying has been applied in mouse [1][2][3][4][5][6][7][8], rat [9,10], rabbit [11], bull [12], boar [13,14] and human [15] spermatozoa. An advantage in freeze-drying is to store the spermatozoa without the use of liquid nitrogen for a long period.…”

Section: Introductionmentioning

confidence: 99%

Pronuclear formation of freeze-dried canine spermatozoa microinjected into mouse oocytes

Watanabe

Asano

Abe

et al. 2009

J Assist Reprod Genet

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“…Intracytoplasmic injection of freeze-dried spermatozoa has been resulted in successful production of live offspring in mouse [1,[4][5][6]32], rat [3,7,33] and rabbit [2], and blastocyst-stage embryos in cattle [8,9] and pig [10]. However, long-term storage of FD spermatozoa at room temperature has not yet been achieved in any mammalian species.…”

Section: Discussionmentioning

confidence: 99%

“…Since the freeze-drying process causes functional losses of sperm cells such as immobilization, the rehydrated spermatozoa need to be injected into oocytes (ICSI: intracytoplasmic sperm injection). Offspring derived from the intracytoplasmic injection of FD spermatozoa have been reported firstly in mouse [1], followed by rabbit [2] and rat [3]. To date, preservation of FD spermatozoa for extended periods (≥1 year) was valid only at a refrigeration temperature in mouse [4, 5, and 6], rabbit [2] and rat [7].…”

Section: Introductionmentioning

confidence: 99%

“…Offspring derived from the intracytoplasmic injection of FD spermatozoa have been reported firstly in mouse [1], followed by rabbit [2] and rat [3]. To date, preservation of FD spermatozoa for extended periods (≥1 year) was valid only at a refrigeration temperature in mouse [4, 5, and 6], rabbit [2] and rat [7]. On the other hand, blastocyst development after ICSI with FD sperm heads was reported in large domestic animals including cattle [8,9] and pig [10].…”

Section: Introductionmentioning

confidence: 99%

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The ability of freeze-dried bull spermatozoa to induce calcium oscillations and resumption of meiosis

2009

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This study was designed to investigate the ability of freeze-dried (FD) bull spermatozoa to induce calcium oscillations in mouse oocytes and meiosis resumption in in-vitro-matured bovine oocytes after intracytoplasmic sperm injection (ICSI). Bull spermatozoa were freeze-dried and stored for one year at +25 ْ◌C, +4 ْ◌C, or -196 ْ◌C. In the first experiment, rehydrated sperm heads were microinseminated into hybrid mouse oocytes loaded with fluo-3/AM, and the kinetics of intracellular calcium concentration was monitored for 1 h. Repetitive increases of intracellular calcium concentration were recorded in the majority of injected oocytes, with exception of a few oocytes injected with FD sperm heads stored at +4 ْ◌C (11%) and +25 ْ◌C (8%) that exhibited a single increase or no response (non-oscillated). Proportion of oocytes oscillated with high frequency (≥10 spikes per hour) was significantly higher (P <0.05) in the non-dried control group (79%) than those in the FD groups (58, 55 and 54% for storage at -196, +4, and +25 ْ◌C, respectively). In the second experiment, control and FD spermatozoa were microinseminated into in-vitro-matured, denuded bovine oocytes. The oocytes were fixed and stained 12 h after ICSI. Significantly higher proportion of bovine oocytes injected with control spermatozoa (70%) resumed meiosis than those injected with +25, +4 and -196 ْ◌C-stored FD spermatozoa (53, 48 and 57%, respectively). The proportion of ICSI oocytes that developed to the pronuclear-stage (complete activation) was significantly higher in the control group (64%) than those in all the FD groups (34, 27, and 28% for storage at -196, +4, and +25 ْ◌C, respectively). Thus, the ability of bull spermatozoa to induce frequent intracellular calcium spikes in mouse oocytes was impaired by the process of freeze-drying, probably resulting in lower proportion of bovine oocytes that resumed meiosis and/or developed to the pronuclear-stage.

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Live rats resulting from injection of oocytes with spermatozoa freeze‐dried and stored for one year

Hochi

Watanabe

Kato

et al. 2007

Molecular Reproduction Devel

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This study was designed to examine whether rat spermatozoa after freeze-drying and 1-year storage can participate in full-term development following intracytoplasmic sperm injection (ICSI). Cauda epididymal spermatozoa from Crlj:Wistar rats were frozen in liquid nitrogen (LN 2 ), first dried for 14 hr at 0.37 hPa and then for 3 hr at 0.001 hPa. The dried spermatozoa were stored for 1 year in a desiccator at þ258C, or in a refrigerator at þ48C, or in LN 2 at À1968C. Controls consisted of sperm that had only been frozen and stored in LN 2 . After being stored, spermatozoa were sonicated to dissociate the sperm tail and were injected into oocytes from superovulated Slc:SD rats. The respective fertilization rates of oocytes injected with frozen sperm, or with freeze-dried sperm stored at þ25, þ4, and À1968C were 79%, 75%, 70%, and 73%. However, the corresponding cleavage rates of injected oocytes were 63%, 1%, 38%, and 36%. After transfer of >80 zygotes of each group into recipients, the respective percentages of full-term normal offspring resulting from frozen sperm or from freeze-dried sperm stored at þ25, þ4, and À1968C were 36%, 0%, 7%, and 14%. These results demonstrate that the storage temperature significantly influenced the likelihood of term development of rats produced by injection of oocytes with freezedried spermatozoa. Chromosomal analysis of the rat spermatozoa in the ICSI oocytes indicated that chromosomal aberration in freeze-dried spermatozoa stored at þ258C (100%) occurred more frequently than in frozen control spermatozoa (41%) and freezedried spermatozoa stored at À1968C (35%), and the frequency of chromosomal aberrations in freezedried spermatozoa stored at þ48C (65%) was the intermediate. In conclusion, rat spermatozoa freezedried and stored at þ48C for 1 year are capable of participating in full-term development after ICSI. Mol. Reprod. Dev. 75: 890-894, 2008.

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Freeze-Dried Sperm Fertilization Leads to Full-Term Development in Rabbits1

Cited by 164 publications

References 31 publications

Pronuclear formation of freeze-dried canine spermatozoa microinjected into mouse oocytes

Pronuclear formation of freeze-dried canine spermatozoa microinjected into mouse oocytes

The ability of freeze-dried bull spermatozoa to induce calcium oscillations and resumption of meiosis

Live rats resulting from injection of oocytes with spermatozoa freeze‐dried and stored for one year

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