• Clinical and Experimental Reproductive Medicine
• /
• v.23 no.3
• /
• pp.303-310
• /
• 1996

The objective of this study was to examine the effect of embryos development following IVF of in vitro-matured porcine oocytes treated with epidermal growth factor (EGF). When cumulus-enclosed oocytes were incubated in TCM 199 medium supplemented with (1) control group, (2) 10 ng/ml EGF, (3) 10${\mu}g$ml FSH and 10% FBS, or (4) 10 ng/ml EGF, 10 ${\mu}g$/ml FSH, and 10% FBS for 42 hr, the late developmental rates on NCSU (0.4% BSA) medium after fertilization were higher in (3) and (4) groups (13.4, 18.3%) than in (2) group (5.2%, p < 0.005), but (2) group is significantly higher than the development to blastocyst of oocytes of (1) group (1.2%). Also, when the cell number of total, ICM, and TE of those blastocysts at 6 day produced in vitro was investigated by double staining (PI and bisbenzimide), total cell number of (4) group (58.80${\pm}$ 11.90) was higher than that of (2) and (3) groups (42.17${\pm}$9.97, 49.07${\pm}$9.77, P < 0.05). ICM cell number of blastocysts of (4) group (11.69${\pm}$5.56) was higher than that of (2) and (3) groups (5.00${\pm}$4.24, 6.77${\pm}$4. 92, P < 0.05). Furthermore, the proportion of ICM in (4) group (19.0${\pm}$1.6) was higher than that in (2) and (3) groups (11.1${\pm}$3.0, 12. 7${\pm}$2.1). These results suggested that in vitromatured porcine oocytes treated with EGF alone can be developed to blastocyst, but high proportion on the development to blastocyst and number of total cell and ICM in blastocyst can be obtained when supplemented with additional FSH and FBS.

• Clinical and Experimental Reproductive Medicine
• /
• v.23 no.3
• /
• pp.319-326
• /
• 1996

The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.

• Korean journal of applied entomology
• /
• v.44 no.4 s.141
• /
• pp.325-330
• /
• 2005

This study was conducted to determine the effect of temperatures on the egg and nymphal development, adult longevity and oviposition of bean bug, Riptortus clavatus Thunberg, using Saealkong seed as food sources in fibrous nylon-tube at four different temperatures (20, 24, 28 and $32^{\circ}C$). Hatchability showed the highest value of 100% at $28^{\circ}C$ and decreased with increasing temperature. Egg duration ranged from 7 days at $32^{\circ}$ to 16.7 days at $20^{\circ}C$. Instar duration was longer with increasing instar stage. Nymphal duration was 38 days at $20^{\circ}C$, 30 days at $24^{\circ}C$, 23 days at $28^{\circ}C$, and 18 days at $32^{\circ}C$ Emergence rates to adult were 16, 41, 72 and 68% at 20, 24, 28 and $32^{\circ}C$, respectively. Female adult longelity ranged from a minimum 20 days at $20^{\circ}C$ to a maximum 63 days at $28^{\circ}C$, while the longevity of male ranged from 19 days at $20^{\circ}C$ to 60 days at $28^{\circ}C$. Preoviposition duration was shorter with increasing temperature and ranged from 11 days at $20^{\circ}C$ to 5 days at $32^{\circ}C$. Total number of eggs laid per female ranged from a minimum 21 eggs at $20^{\circ}C$ to a maximum 67 eggs at $28^{\circ}C$. Consequently, the estimated lower threshold tempeatures of each developmental stage were $10.3^{\circ}C$ for egg, and 9.3, 12.7, 10.0, 11.0 and 8.7 for the 1st, 2nd, 3rd, 4th and 5th instar, respectively.

• Korean journal of applied entomology
• /
• v.44 no.4 s.141
• /
• pp.277-282
• /
• 2005

This study was conducted to investigate morphological characteristics and effects of temperature on the development of Piezodorus hybneri on soybean. The unibanded stink bug, Piezodorus hybneri, is a serious insect pest in soybean fields giving damage to seeds in pod and leaves of soybean lowering both quality and yield. Eggs were spherical and laid in two raws on the leaves and pods of soybean plants. Body lengths of females and males were 9.8 mm and 8.7 mm, respectively. Egg hatch rates were better in higher temperature within the range of examined temperatures, which ranged in $81.2{sim}93.2%$. The development periods of eggs at the temperatures of 20, 25, 30 and $35^{\circ}C$ were 10.7, 5.0, 4.0, and 3.0 days, respectively. Mean developmental periods of 1st, 2nd, 3rd, 4th and 5th nymphs at $25^{\circ}C$ were 3.2, 3.4, 3.4, 3.3 and 5.9 days, respectively. Development threshold and effective accumulative temperature were $13.3^{\circ}C$ and 65.5 DD (day degree) for egg stage, $9.9^{\circ}C$ and 322.8 DD for nymph stage, $10.7^{\circ}C$ and 386.4 DD from egg to adult, respectively. Oviposition began from 10 days after emergence at $25^{\circ}C$, and the longevity of female and male were 52.5 and 38.2 days, respectively. Total number of eggs and egg masses laid by a female at $25^{\circ}C$ were 496 and 21.3, respectively, The longevity of adult female was shortened with increasing temperature, whereas the total numbers of eggs laid by a female were decreased.

• Development and Reproduction
• /
• v.13 no.1
• /
• pp.1-11
• /
• 2009

Implantation itself is governed by an array of endocrine, paracrine and autocrine modulators, of embryonic and maternal origin. Window of implantation is the unique temporal and spatial expression of factors allows the embryo to implant via signaling, appositioning, attachment, and invasion in a specific time frame of $2{\sim}4$ days. When the embryo has arrived in the uterine cavity, a preprogrammed sequence of events occurs, which involves the production and secretion of a multitude of biochemical factors such as cytokines, growth factors, and adhesion molecules by the endometrium and the embryo, thus leading to the formation of a receptive endometrium. Cytokines such as LIF, CSF-1, and IL-1 have all been shown to play important roles in the cascade of events that leads to implantation. Integrin, L-selectin ligands, glycodelin, mucin-1, HB-EGF and pinopodes are involved in appositioning and attachment. The embryo also produces cytokines and growth factors (ILs, VEGF) and receptors for endometrial signals such as LIF, CSF-1, IGF and HB-EGF. The immune system and angiogenesis play an important role. The usefulness of these factors to assess endometrial receptivity and to estimate the prognosis for pregnancy in natural and artificial cycles remains to be proven. Integrins, pinopodes, glycodelin and LIF (from biopsies) are promising candidates; from uterine flushings, glycodelin and LIF are also candidates. The ideal serum marker is not available, but VEGF, glycodelin and CSF have some clinical implications. Further evaluation that includes larger groups of infertile women and fertile controls are needed to elucidate whether their presence in plasma, flushing fluid, or endometrial samples can be used as some kind of a screening tool to assess endometrial function and prognosis for pregnancy before and after artificial reproductive therapy. A better understanding of their function in human implantation may lead to therapeutic intervention, thereby improving the success rate in reproduction treatment. New molecular techniques are becoming available for measuring both embryonic and endometrial changes prior to and during implantation. The use of predictive sets of markers may prove to be more reliable than a single marker. Ultimately, the aim is to use these tools to increase implantation in artificial cycles and consequently improve live-birth rates.

• Korean Journal of Animal Reproduction
• /
• v.25 no.1
• /
• pp.1-7
• /
• 2001

This study was to test whether in vitro matured Hanwoo oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5~10 min, exposed in EG30 for 30 sec, each oocyte was individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures 〔1.0 M sucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS〕 at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were pregnant and three of them were ongoing-pregnant by manual palpation at 250 days after transfer. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

• Journal of Embryo Transfer
• /
• v.26 no.1
• /
• pp.19-25
• /
• 2011

This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.