• Clinical and Experimental Reproductive Medicine
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• v.31 no.4
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• pp.209-215
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• 2004

Objective: Embryonic stem (ES) cells could be differentiated into the specific cell types by alternation of culture condition and modification of gene expression. This study was performed to evaluate the differentiation protocol for mouse and human ES cells to insulin secreting cells. Methods: Undifferentiated mouse (JH-I) and human (Miz-hESI) ES cells were cultured on STO feeder layer, and embryoid bodies (EBs) were formed by suspension culture. For the differentiation, EBs were cultured by sequential system with three stage protocol. The differentiating ES cells were collected and marker gene expressions were analyzed by seIni-quantitative RT-PCR in each stage. Amount of secreted insulin levels in culture media of human ES cells were measured by human insulin specific RIA kit. Results: During the differentiation process of human ES cells, GATA-4, a-fetoprotein, glucose transporter-2 and Ngn-3 expression were increased whereas OctA was decreased progressively. Insulin and albuInin mRNAs were expressed from stage IT in mouse ES cells and from stage III in human ES cells. We detected 3.0~7.9 IlU/rnl secretion of insulin from differentiated human ES cells by in vitro culture for 36 days. Conclusion: The sequential culture system could induce the differentiation of mouse and human ES cells into insulin secreting cells. This is the fIrst report of differentiation of human ES cells into insulin secreting cells by in vitro culture with serum and insulin free medium.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.4
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• pp.203-209
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• 2011

Objective: This study was performed to compare the efficiency of slow freezing and vitrification based on survival, development to blastocysts, and cell numbers of blastocysts. Changes in embryonic gene expression in fresh and frozen-thawed embryos were also examined. Methods: Eight-cell stage embryos were collected from superovulated female BDF1 mice. The collected embryos were randomly divided into three groups. One group was maintained as fresh controls (n=42), one was frozen by slow freezing (n=43), and one was cooled by vitrification (n=43). After thawing or cooling, survival rates, development to blastocyst, and cell numbers and inner cell mass (ICM) cell numbers of blastocysts were compared with those of the control group. The expressions of eight genes ($Rbm3$, $Birc5$, $Sod1$, $Sod2$, $Cirbp$, $Caspase3$, $Trp53$, $Hsp70.1$) were examined by real time-quantitative polymerase chain reaction in the fresh and frozen-thawed embryos. Results: There were no significant differences in the slow freezing and vitrification groups' survival rate after thawing (88.4% vs. 88.4%), development to blastocyst (100% vs. 97.4%), cell numbers ($107.0{\pm}21.0$ vs. $115.0{\pm}19.7$), or ICM cell numbers of blastocysts ($11.3{\pm}5.2$ vs. $11.1{\pm}3.7$). Cell numbers of blastocysts were significantly ($p$ <0.05) lower in the frozen-thawed embryos than the fresh embryos. There were no significant differences in the slow freezing and the vitrification groups' expressions of the eight genes. The expressions of $CirbP$ and $Hsp70.1$ were higher in the frozen-thawed embryos than in the fresh embryos but there were no significant differences. Conclusion: These results suggest that there were no significant differences between embryos that underwent slow freezing and vitrification.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.2
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• pp.45-50
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• 2015

Objective: Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. Methods: The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n=480; AOA, n=29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n=12) and non-motile sperm-injected (n=17) groups (total n=29 cycles). Results: The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or delivery rate (5.9% vs. 16.7%) between the two groups. Conclusion: We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure.

• Clinical and Experimental Reproductive Medicine
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• v.39 no.4
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• pp.144-152
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• 2012

Objective: Concerns are growing about the decrease in male reproductive health. Caffeine is one of the popular nutrients that has been implicated as a risk factor for infertility. In the present study, we examined whether in utero and lactational exposure to caffeine affects the reproductive function of the offspring of rats. Methods: Pregnant rats received caffeine via drinking water during gestation (26 and 45 mg/kg) and lactation (25 and 35 mg/kg). Body and reproductive organ weight, seminiferous tubule diameter, germinal epithelium height, sperm parameters, fertility rate, number of implantations, and testosterone level of the offspring were assessed from birth to adulthood. Results: Significant dose-related decreases were observed in the body and reproductive organ weight, seminiferous tubule diameter, and germinal epithelium height of the offspring. Sperm density had declined significantly in offspring of the low-dose and high-dose groups, by 8.81% and 19.97%, respectively, by postnatal day 150. The number of viable fetuses had decreased significantly in females mated with male offspring of the high-dose group at postnatal days 60, 90, 120, and 150. There were also significant reductions in testosterone levels of high-dose group offspring from birth to postnatal day 150. Conclusion: It is concluded that maternal caffeine consumption impairs gonadal development and has long-term adverse effects on the reproductive efficiency of male offspring rats.

• Advances in Traditional Medicine
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• v.4 no.2
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• pp.77-81
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• 2004

Petroleum ether, benzene and alcohol extracts of seeds of C. juncea administered orally at the dose level of 25mg/100g body weight to adult female mice for 30 days, resulted in irregular estrous cycle with prolonged estrus and metaestrus and reduced diestrus and proestrus during the experimental period. Histological studies of the ovary indicate increases in the number of atretic follicles but decreases in the number of developing follicles, Graafian follicles and corpora lutea. The total cholesterol content of the ovary is increased, whereas ascorbic acid content is decreased. The weight of the uterus and its micrometric measurement in all experimental mice are increased significantly. The alcoholic extracts showed estrogenic activity in immature mice by early opening of the vagina, premature cornification of the vaginal epithilium and increases in uterine weight. However, alcohol extract of seeds of C. juncea was more effective in causing these changes compared to other extracts. After subjecting to preliminary phytochemical screenings alcohol extract showed positive; test for alkaloids, steroids, glycosides, flavones, phenols and tannins.

• Animal cells and systems
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• v.5 no.2
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• pp.127-132
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• 2001

The object of this study is to differentiate the reproductive behaviors of Pungitius sinensis and P. kaibarae inhabiting the southern part of the Korean Peninsula. We collected P. sinensis from Jusu stream, Okkyemyeon and P. kaibarae from Sacheon stream, Sacheon-myeon both in Gangwon province and subsequently raised and observed them in an aquarium. At the beginning of the reproductive season, male P. sinensis got tinged with dark green, made a territory, and built nests on the bottom. On the other hand, male P. kaibarae became black all over, its white ventral spines became conspicuous and built nests on the stems of waterweed off the bottom. In the courtship dance, male P. sinensis made frequent attempts to entice females into their nests after many bitings, while male P. kaibarae mostly did this with conspicuous jumpings. In courtship behaviors, the body's angle of male P. kaibarae with his head down was larger than that of male P. sinensis by 50-60 degrees. During courtship, the biting frequency as an index of aggressive behavior was greater in P. sinensis and the jumping frequency as an index of sexual behavior was greater in P. kaibarae. During the courtship dance, bitings tended to suppress jumpings, for P. sinensis, but not for P. kaibarae.

• Applied Microscopy
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• v.43 no.3
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• pp.103-109
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• 2013

ICI 182,780 (ICI) is known as an estrogen receptor antagonist, whereas propyl pyrazole triol (PPT) is an estrogen receptor agonist. In this study, ICI or ICI added with PPT was injected into adult male mice. Body and reproductive organ weights were reduced in the ICI added with PPT group compared to the control group. Further, the ICI and ICI added with PPT groups both showed increases in luminal areas of the seminiferous tubules of the testis, whereas cell heights of efferent ductules and the initial segment of the epididymis were reduced. Sperm count in the caudal epididymis was reduced in the ICI and ICI added with PPT groups. These results show that reproductive tissues were more deeply affected in the ICI added with PPT group. We also demonstrated that treatment with ICI resulted in histological changes in the testis, efferent ductule, and epididymis. Further, alternate treatment with ICI and PPT induced abnormalities in reproductive organs. These results indicate that a high concentration of PPT together with ICI may cause histological abnormalities instead of histological restoration in reproductive organs.

• Development and Reproduction
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• v.19 no.4
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• pp.167-180
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• 2015

Arsenic is a toxic metalloid that exists ubiquitously in the environment, and affects global health problems due to its carcinogenicity. In most populations, the main source of arsenic exposure is the drinking water. In drinking water, chronic exposure to arsenic is associated with increased risks of various cancers including those of skin, lung, bladder, and liver, as well as numerous other non-cancer diseases including gastrointestinal and cardiovascular diseases, diabetes, and neurologic and cognitive problems. Recent emerging evidences suggest that arsenic exposure affects the reproductive and developmental toxicity. Prenatal exposure to inorganic arsenic causes adverse pregnancy outcomes and children's health problems. Some epidemiological studies have reported that arsenic exposure induces premature delivery, spontaneous abortion, and stillbirth. In animal studies, inorganic arsenic also causes fetal malformation, growth retardation, and fetal death. These toxic effects depend on dose, route and gestation periods of arsenic exposure. In males, inorganic arsenic causes reproductive dysfunctions including reductions of the testis weights, accessory sex organs weights, and epididymal sperm counts. In addition, inorganic arsenic exposure also induces alterations of spermatogenesis, reductions of testosterone and gonadotrophins, and disruptions of steroidogenesis. However, the reproductive and developmental problems following arsenic exposure are poorly understood, and the molecular mechanism of arsenic-induced reproductive toxicity remains unclear. Thus, we further investigated several possible mechanisms underlying arsenic-induced reproductive toxicity.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.371-380
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• 1999

Animal reproductive performance could be affected by several regulatory factors, including nutritional, environmental, and genetic factors. Particularly, during the last half of this century, animal reproductive performance has been remarkably successful in improving the efficiency of livestock production. For some traits efficiency gains have been achieved with little or no knowledge of the genes underlying the traits. And, they have depended upon the phenotypic selection by statistical methods to estimate the genetic parameters of some reproductive traits. In spite of these successes, it is clear that recent advances in both developmental biology and molecular biology are set to revolutionize he practice of animal reproductive performance n the 21th century. (omitted)

• 대한생식의학회:학술대회논문집
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• 2006.11a
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• pp.35-36
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• 2006