Principles of the modern bioethics and the ethical status of the fetus are interpreted in this article. The modern bioethics is based on the three basic ethical principles reported in Belmont report. The principles consist of respect for persons, beneficence and justice. Respect for persons was considered to be the most important principle and have the priority among the principles. However some ethicist dissent from the priority of respect for persons. When we face a case in which the principles conflict each other, the top priority needs to be granted to one of the principles. How we select one of the principles for the top priority remains an unsolved issue. Considering the ethical status of the fetus, the most important thing is when the embryo or fetus becomes a person. Roman Catholic declared in 1974 that the embryo just after fertilization is considered as a person. However many bio-ethicists believe that only an autonomous person who is an individual capable of deliberation about personal goals, and of acting under the direction of such deliberation has the right of living. This opinion permits us not only to abort fetus artificially but also to kill infants. In the latter half of the article personal opinion on the ethical status of the fetus is explained.

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It has been a problem that the rate of development of porcine embryos generated by intracytoplasmic sperm injection (ICSI) is quite low. In this issue, we introduce ideas from recent studies to improve the developmental competence of porcine ICSI embryos: uniqueness in physiology of fertilization by ICSI, and the effects of sperm preparation before ICSI on completion of fertilization and embryonic development following ICSI.

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Chromosomal abnormalities were evaluated in mouse embryos derived from in vitro fertilization using oocytes from old (12 months) BALB/c (12-BALB) and ICR (12-ICR) females and young (3 months; 3-BALB and 3-ICR) female mice. Chromosomal analysis was carried out at two developmental stages, 12-BALB and 12-ICR vs 3-BALB and 3-ICR at the first cleavage division and 12-BALB vs 3-ICR at the second division. Although all young females ovulated, this was not so in the old females; additionally, significantly fewer oocytes were ovulated by the old females and the fertilization rate was significantly lower in the 12-ICR group, but not in the 12-BALB group. Significantly fewer of the embryos in the old group had a metaphase figure at the first cleavage division and this correlated with significantly more of the embryos in this group being still at the pronuclear stage. The incidence of polyploidy, probably due to polyspermy, was found in both the 12-BALB and 12-ICR groups at the first cleavage division, but this was not seen in embryos at the second division. These results indicate that the aging of females impairs reproductive success, as evidenced by anovulation, polyspermy, and delays in fertilization/asynchronous development in resulting embryos.

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We investigated the indication of primary ICSI. In 639 cycles (356 cases) of conventional IVF, we examined the fertilization rate and pregnancy rate upon following items. Those were appearance and concentration of sperm, female age, and picked up oocyte. In the study of correlation between sperm concentration and rate of fertilization or pregnancy, the cases of sperm concentration less than 10 × 10⁶/ml were fertilized only 13.3% (18/135) or no cases were got pregnant (0/21). But in the study of correlation between sperm motility rate and rate of fertilization or pregnancy, even in cases of motility rate less than 20%, fertilization rate was 33% (43/130) and pregnant rate was 19% (4/21). We also considered the sperm concentration after the swim up cases. In the cases sperm concentration less than 1 × 10 ⁶/ml, fertilization rate was only 2.3% (2/86) or no cases got pregnant (0/13). We could not found definite correlation between female ages or quantity of picked up oocyte and rate of fertilization and pregnancy respectively. It mentioned that these factors are not applied to the indication of primary ICSI. We concluded that the indication of primary ICSI should be based on the sperm concentration and the numerical numbers were less than 10 × 10 ⁶/ml for ordinary cases and less than 1 × 10 ⁶/ml for after the swim up cases.

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The indication of ICSI is generally based on sperm analysis, past therapeutic history, and prediction of fertilization failure. In this study, we attempted to evaluate the usefulness of SMI (Sperm Motility Index) and "split cycle" procedure on ICSI indication. According to SMI score, patients were separated into three groups: I) Poor (0≤SMI<80, n=10), II) Medium (80≤SMI<160, n=9), III) Good (160≤SMI, n=43). Among these, no significant difference was observed in fertilization rates (Poor 56.4%, Medium 69.6%, and Good 71.1%) or in pregnancy rates (20.0%, 33.3%, and 48.8%, respectively). On the other hand, patients were separated into three groups: I) conventional IVF (n=188), II) split cycle (n=14), III) ICSI (n=72). Among these, no significant difference was observed in fertilization rates (conventional IVF 64.8%, split cycle 63.0%, and ICSI 56.0%) or in pregnancy rates (46.6%, 57.1%, and 31.7%, respectively). Surprisingly, among those in split cycle who had had fertilization failure on previous conventional IVF trial, no significant difference was found in fertilization rates between conventional IVF eggs and ICSI eggs. In such cases, split cycle might be a better choice.

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The purpose of this study is to determine criteria of split ICSI in which half of oocytes were inseminated by ICSI and half by conventional IVF. Six hundreds eighty-two couples who experienced the first assisted reproductive technology in our clinic were enrolled in the study. Pregnancy rate in couples with fertilization rate (FR) less than 30% was significantly lower than that in couples with FR exceeding 50%. FR in couples with oligozoospermia (sperm count < 20 × 10⁶/ml) (50.0~53.8%) was significantly lower than that in couples with normozoospermia (65.0~79.5%). FR in couples with sperm motility rate less than 20% (0~29.6%) were significantly lower than that in couples with motility rate exceeding 20% (66.8~76.8%). Infertile couples with oligozoospermic semen or low sperm motility rate (<20%) should be treated by split ICSI rather than by IVF.

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The cycles were divided into the following five groups (100-500, 500-1,000, 1,000-1,500, 1,500-2,000, ≥2,000 × 10⁴/ml) by the concentration of motile sperm and compared the rates of normal fertilization and cycle from which normal fertilization was not obtained. In motile sperm concentration from 100 to <500 × 10⁴/ml, the normal fertilization rate was significantly lower (48.8%) than other groups. The accordance with the aging and declining sperm counts, the normal fertilization rate has declined and ration of the cycle, which could not obtain normal fertilization has risen. This study suggested that the cycle in motile sperm concentration from 100 to <500 × 10⁴/ml is not be suitable for IVF. Moreover, the cycle in motile sperm concentration from 500 to <1,000 × 10⁴/ml, we have to consider the number of oocyte, female age and number of treatment cycle.

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The blastocyst transfer (BT), transferring blastocysts following IVF, has been recently focused as an effective assisted reproductive technology in humans because it has powerful efficacy on not only prevention of multiple pregnancies but also high pregnancy rates. The BT has been already applied to about 70% in IVF cycles per year in our facility resulted in excellent clinical results. Quality of the resulting blastocyst is the most important key for the successful BT. High quality blastocysts can be produced in vitro by the sequential culture method using commercially supplied media under low oxygen atmosphere phase in a multi-gas incubator. Optimized environments of the embryo culture room (27°C, high humidity and bio-clean room without ultraviolet light) and completed conditions of the culture media are essential for production of the excellent blastocysts. Other fine advices for success are also introduced in the present article.

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Conditions of optical microscopes and ways to use them are directly related to the easiness or difficulty of ICSI procedures. The attached instruction book explains how to use and maintain the microscope in detail. In terms of an optics maker, perusing the instructions are strongly recommend. However, some users sometimes claim that they feel difficulty to understand an instruction. In this paper how to use and maintain the microscopes used in ICSI-for example a stereoscopic microscope, a phase contrast microscope, an inverted microscope fitted with Nomarski differential interference optics and an inverted microscope with Hoffman optics-is plainly explained with a lot of figures.

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Wrong:English abstract of this article was left out.
Right:Abstract: It is essential for successful ICSI to select the spermatozoa with good motility and morphology and keep the quality of gametes throughout ICSI procedure. Embryogists are required a fundamental knowledge of the gametes for protection of them. Especially, we must take grate care to treat cumulus oocyte complexes with hyaluronidase solution and immobilize motile spermatozoa and inject a spermatozoon into oocyte.
Key words: ICSI, Immobilization of spermatozoa, Polyvinyl pyrrolidone, Spindle

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