Fertilization of Chinese Hamster Eggs in Vitro

Pickworth, Stella; Chang, M. C.

doi:10.1530/jrf.0.0190371

Cited by 35 publications

(12 citation statements)

References 7 publications

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“…As far as we are aware, successful in vitro fertilization in the Chinese hamster has been reported only by Pickworth and Chang [1969]. They preincubated vas deferens spermatozoa for 2-3 h in media (either Tyrode's solution or Ham F-I0 medium with 1% bovine serum albumin) containing cumulus masses of the golden hamster, then mixed the spermatozoa with Chinese hamster eggs in cumulus.…”

Section: Discussionmentioning

confidence: 99%

Gametes and fertilization in the Chinese hamster

et al. 1983

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Freshly ovulated eggs are each surrounded by a compact cumulus oophorus. The overall diameter of the normal egg (including the zona pellucida) is about 100 μm. Cumulus cells, particularly those near the egg, are arranged redially in a viscous noncellular matrix. The spermatozoon is about 250 μm in length. The head a large acrosome, changes in which can be readily examined with the light (phase‐ contrast) microsope. When exposed to physiological salt solutions, testicular spermatozoa either were motionless or flexed the posterior half of their tails slowly. Spermatozoa from the caput epididymis were highly motile, flexing the entire tail. A few of them moved progressively. Mature spermatozoa from the vas deferens were highly motile and moved either straightforward or in a circle. They vibrated their tails stiffly without flexing them. In normally mated females, fertilization began sometime between 2 and 3 h after ovulation and was completed within the next 4 to 5 h. Spermatozoa swimming in the ampullary fluid or within the cumulus oophorus about the time of fertilization flexed the anterior half (which roughly corresponds to the midpieac region) of their tails. This peculiar movement may be homologous to the so‐called “hyperactivation” of spermatozoa as reported in several other mammalian species. Actively motile spermatozoa within the cumulus or no the zona pellucida had either modified (“collapsed”) or no acrosomal caps. The sperm head usually passed verticually or nearly through the zona, but the path was oblique in some instances. In 54% of the recently fertilized eggs examined, the entire length of the sperm tail was within the perivitelline space; in the other 46% of the eggs varying lenghts of the tail remined the perivitelline space, the tails were extruded from the vitellus of many eggs even before the eggs began their first cleavage. When unfertilized eggs in the cumulus oophorus were inseminated with vas deferens spermatozoa in a modified Tyrode's solution (m‐TALP), about 80% of them were ferrtilized by 4–6 h after insemination. The vast majority were monospermic. When eggs were freed from the cumulus prior to insemination, none were fertilized, suggesting that the cumulus cells or their matrix assisted capacitation and/or the acrosome reaction of the spermatozoa under the in vitro conditions employed. No eggs were fertilized by the testicular or caput epididymal spermatozoa regardless of the presence or absence of cumulus oophorus around the eggs at the time of insemination.

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Section: Discussionmentioning

confidence: 99%

Gametes and fertilization in the Chinese hamster

et al. 1983

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“…However, successful techniques for IVF and embryo culture have not been developed in murine species except for the mouse (Kasai et al, 1978;Quinn & Whirtingham, 1982;Chatot et al, 1990;Summers et al, 1995) and rat (Miyoshi et al, 1995a, b). With respect to the Chinese hamster, some investigators have reported methods for IVF (Pickworth & Chang, 1969;Yanagimachi et al, 1983;Parkening & Cisneros, 1988) and in vitro culture of embryos (Basler & Rohrborn, 1977;Parkening & Cisneros, 1988). However, no-one has yet succeeded in the in vitro culture of fertilised ova All correspondence to: Dr H. Tateno, Department of Biological Sciences, Asahikawa Medical College, 4-5 Nishikagura, Asahikawa 078, Japan.…”

Section: Introductionmentioning

confidence: 99%

In vitrofertilisation of Chinese hamster oocytes by spermatozoa that have undergone ionophore A23187-induced acrosome reaction, and their subsequent development into blastocysts

Tateno

Kamiguchi

1996

Zygote

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SummaryTo enhance potential use of the Chinese hamster, Cricetulus griseus, in developmental and cytogenetic studies of mammalian gametes and embryos, techniques for in vitro fertilisation and embryo culture were developed in the species. Spermatozoa were recovered from the vasa deferentia of mature males, and incubated in modified TYH medium for 1 h at 37°C under 5% CO2 in air. They were then treated with ionophore A23187 (20¼M) for 10min to induce the acrosome reaction. Following ionophore treatment, superovulated oocytes were collected from hormonally stimulated females and incubated with the acrosome-reacted spermatozoa for 2 h at 37°C under 5% CO2 in air. In this study, 245 oocytes ova (98.0%) were determined to be monospermic. The monospermic ova were then cultured in TYH supplemented with 1mM hypotaurine under the same gas phase. Within 30h of fertilisation, 182 ova (93.8%) cleaved to the 2-cell stage, and subsequently 163 ova (84.0%) developed beyond the 2-cell stage. Thus, obstinate developmental arrest at the 2-cell stage(‘2-cell block’) was not observed in this species. Ultimately, 65.5% of monospermic ova reached morula to blastocyst stages.

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“…The Chinese hamster, Cricetulus griseus, introduced as a laboratory animal more than 60 years ago [Yerganian, 19581, became attractive for reproductive studies only in recent years [Pickworth et al, 1968;Pickworth and Chang, 1969;Yanagimachi et al, 1983, 19851. Pickworth andChang [1969] were the first to report that ovulation can be induced in both immature and maturing Chinese hamsters by treating females with PMSG (5 IU) and hCG (10 IU) 48-52 hr apart. However, it is not clear how many superovulated eggs they could obtain.…”

Section: Introductionmentioning

confidence: 99%

Superovulation in immature and mature Chinese hamsters

1987

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Mature female Chinese hamsters ovulate an average of 8.8 +/- 1.0 (mean +/- SD) eggs per female in each estrous cycle. Superovulation can be induced in both immature and mature females by subcutaneous or intraperitoneal injections of pregnant mare serum gonadotropin (PMSG) and either human chorionic gonadotropin (hCG) or pituitary luteinizing hormone (PLH). The best superovulation in immature females was induced by the administration of 15 IU of PMSG followed 72 hr later by injection of 15 IU of hCG (about 25 eggs per female) or 0.2 mg (200 IU) PLH (about 46 eggs per female). Ovulation started about 13-15 hr after administration of hCG (or PLH) and was completed during the next 5-6 hr. Superovulation in mature females could be induced by injecting PMSG any day of the estrous cycle, but the best superovulation (about 39 eggs per female) was induced by injecting 15 IU of PMSG on day 1 (day of ovulation) followed by the injection of 0.4 mg of PLH 72 hr later. When immature females treated with the best superovulatory protocol were mated on the evening of PLH injection, only 5% of the eggs were found fertilized 50 hr after PLH administration. On the other hand, about 60% of the eggs were found fertilized in mature females mated following treatment with the best superovulatory protocol. The majority (83-85%) of superovulated eggs obtained from both immature and mature females were normally fertilized in vitro.

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Fertilization of Chinese Hamster Eggs in Vitro

Cited by 35 publications

References 7 publications

Gametes and fertilization in the Chinese hamster

Gametes and fertilization in the Chinese hamster

In vitrofertilisation of Chinese hamster oocytes by spermatozoa that have undergone ionophore A23187-induced acrosome reaction, and their subsequent development into blastocysts

Superovulation in immature and mature Chinese hamsters

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