• Journal of Embryo Transfer
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• v.25 no.1
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• pp.9-14
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• 2010

This study was conducted to find out the effects of artificial shrinkage (AS) on post-thaw development of bovine embryos. The blastocoelic cavity of blastocyst was punctured to remove its fluid contents and then incubated in the holding medium (HM) for 10 min. The punctured and non-punctured (control) blastocysts were equilibrated in vitrification solution 1 (VS1; TCM-199+20% FBS+10% EG) for 5 min and vitrification solution 2 (VS2; TCM199+20% FBS+35% EG+5% PVP+0.5 M Sucrose) for 1 min and vitrified by direct dropping into the liquid nitrogen. Vitrified blastocysts (punctured and control) were thawed and cultured in vitro (12 hr) for studying survival and hatching rates. The levels of shrinkage were measured by the volume of the blastocyst during equilibration in VS1 (at 1, 3 and 5 min of equilibration) and VS2 (at 30 and 60 sec of equilibration) that was considering the volume of non-punctured blastocyst in HM as 100%. The levels of shrinkage were higher in punctured group (62.4, 64.6, 64.3% at 1, 3 and 5 min in VS1; 50.6 and 52.7% at 30 and 60 sec in VS2) than control group (84.8, 86.6, 86.4% at 1, 3 and 5 min in VS1; 72.1 and 68.8% at 30 and 60 sec in VS2), but within each group the levels of shrinkage were similar. The survival (90.9%) and hatching (50.0%) rates of vitrified blastocysts at 12 hr post-thaw were higher in punctured group than that in control group (76.9% and 0.0% respectively). We confirmed that vitrification solutions (VS1 and VS2) have no toxic effect on the survival of blastocysts because the survival rates of blastocysts exposed to VS1 and VS2 for 24 hr were similar between punctured and control groups (94.3 vs. 96.0%; p>0.05). In conclusion, the preliminary data show that AS of blastocyst may improve survival and hatching rate after thawing.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.1
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• pp.1-7
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• 2015

Stress coping mechanisms are critical to minimize or overcome damage caused by ever changing environmental conditions. They are designed to promote cell survival. The unfolded protein response (UPR) pathway is mobilized in response to the accumulation of unfolded proteins, ultimately in order to regain endoplasmic reticulum (ER) homeostasis. Various elements of coping responses to ER stress including Perk, Ask1, Bip, Chop, Gadd34, Ire1, Atf4, Atf6, and Xbp1 have been identified and were found to be inducible in oocytes and preimplantation embryos, suggesting that, as a normal part of the cellular adaptive mechanism, these coping responses, including the UPR, play a pivotal role in the development of preimplantation embryos. As such, the UPR-associated molecules and pathways may become useful markers for the potential diagnosis of stress conditions for preimplantation embryos. After implantation, ER stress-induced coping responses become physiologically important for a normal decidual response, placentation, and early organogenesis. Attenuation of ER stress coping responses by tauroursodeoxycholate and salubrinal was effective for prevention of cell death of cultured embryos. Further elucidation of new and relevant ER stress coping responses in periimplantation embryos might contribute to a comprehensive understanding of the regulation of normal development of embryonic development and potentiation of embryonic development in vitro.

• Journal of Plant Biotechnology
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• v.31 no.1
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• pp.61-65
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• 2004

In order to micropropagate uniform plantlets of Alocasia cadieri Chantrier in vitro, the shoot tips were cultured on media containing various concentrations of BA and thidiazuron (TDZ). Multiple shoot formation from shoot tips was very effective on medium containing 0.1mg/L TDZ. The formed shoots from shoot tips were separated into a shoot, and cultured on media with BA, TDZ, and NM combination for proliferation. The shoots were multiplied very vigorously on medium with 0.5mg/L TDZ and 0.5mg/L NAA. The rooting and growth of multiplied shoots were more effective on medium with 2.0g/L activated charcoal, rather than those with IBA and NAA. Rooted plantlets show high survival in soil mixed with perlite 1: vermiculite 1 or vermiculite alone.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.123-128
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• 2005

This study examines the effects of kinds and concentrations of cryoprotectants on the survival rate of vitrification-thawed porcine oocytes, together with the effects on survival, in vitro fertilization and development of immature oocytes. 1. The developmental rate of oocytes to MII and diploid stage when the vitrification-thawed of recovered immature oocytes cultured for 0, 15, 30 and 40h were cultured for 0, 15, 30 and 40h were $56.7\%,\;53.3\%,\;63.3\%,\;65.0\%\;and\;23.3\%,\;18.3\%,\;10.0\%,\; 3.3\%$, respectively. The in vitro development to MII stage were lower than the control group $(78.2\%)$, but higher fo. diploid stage $(5.5\%)$. 2. When the vitrification of immature oocytes after being culture for 0, 15, 30 and 40 hours, the survival rate were $34.0\%,\;26.0\%,\;18.0\%\;and\;10.0\%$ respectively. This result was lower than that of the control group $(60.0\%)$. 3. When the fertilization of the vitrified immature oocytes after being culture for 0, 15, 30 and 40 hours, the in vitro fertilization rate were $60.0\%,\;54.0\%,\;48.0\%,\;38.0\%$, and developmental rates were $26.0\%,\;18.0\%,\;8.0\%,\;4.0\%$, respectively. This results were lower than the control group $(78.0\%\;and\;38.0\%)$. 4. When the fertilization of the immature oocytes after being culture for $0\~15$ hours vitrified with EDS and ETS, the fertilization and developmental rates were $50.0\%,\; 22.0\%$ and $46.0\%,\;18.0\%$, respectively. This results were lower than the control group $(74.0\%\;and\;38.0\%)$.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.585-592
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• 2017

Objective: The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods: Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results: The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture ($143.8{\pm}10.5$ to $159.2{\pm}14.8$) was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT) groups ($31.4{\pm}8.3$ to $33.4{\pm}11.1$). After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p<0.05) between sexes. Conclusion: The present study found that the sex difference of the nuclear donor does not affect the developmental rate of porcine SCNT embryos. Furthermore, postnatal survivability of the cloned piglets was not affected by the sex of the donor cell.

• Toxicological Research
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• v.33 no.4
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• pp.273-282
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• 2017

Chemoprevention entails the use of synthetic agents or naturally occurring dietary phytochemicals to prevent cancer development and progression. One promising chemopreventive agent, procyanidin, is a naturally occurring polyphenol that exhibits beneficial health effects including anti-inflammatory, antiproliferative, and antitumor activities. Currently, many preclinical reports suggest procyanidin as a promising lead compound for cancer prevention and treatment. As a potential anticancer agent, procyanidin has been shown to inhibit the proliferation of various cancer cells in "in vitro and in vivo". Procyanidin has numerous targets, many of which are components of intracellular signaling pathways, including proinflammatory mediators, regulators of cell survival and apoptosis, and angiogenic and metastatic mediators, and modulates a set of upstream kinases, transcription factors, and their regulators. Although remarkable progress characterizing the molecular mechanisms and targets underlying the anticancer properties of procyanidin has been made in the past decade, the chemopreventive targets or biomarkers of procyanidin action have not been completely elucidated. This review focuses on the apoptosis and tumor inhibitory effects of procyanidin with respect to its bioavailability.

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

• Korean Journal of Plant Resources
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• v.6 no.2
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• pp.171-179
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• 1993

This study was conducted to investigate the effect of growth regulators and medium composition on the growth of each stage in apical meristem culture for mass propagation of Zanthoxylum piperitum var. inerme Makino. The source material, shoot tip segments were taken from three-years old graft trees. Apical meristems were cultured in vitro on basal MS, GD, WS, half strength MS(1/2MS) and half strength GD(1/2GD) media supplemented with various concentrations for growth regulators(BA, IBA) and inorganic nutrients. The results summarized are as follows: 1. In culture establishment stage, ratio of culture establishment was 96.7% and the best resuit was obtained using MS medium supplemented with 1.0mg/l BA and 0.2mg/l IBA. 2. In shoot multitication stage, both shoot multiplication and growth were achieved in average 5.6cm. These results were obtained on in MS medium supplemented with 1.0mg/l BA and 0.2mg/l IBA. 3. In roothing stage, phloroglucinol(PG) acted as IBA synergist in root initiation. The most faverable combinations for root development was half-strength MS medium supplemented with 162mg/l PG and 0.2mg/l IBA, and ratio of rooting was 58.0%. 4. In Vitro formed plantlets were transplanted to paper pots in greenhouse with 85% of relative humidity. 96% of survival rate was obtained from artificial soil mix having same volume of sand, vermiculite, peat, and soil.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.84-84
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• 2002

The advantages of the OPS techniques(Vajta G et al, Mol Reprod Dev 51: 53-58,1998) give 1) high survival rates of various types of eggs, 2) quick and simple process, 3) inexpensive equipment and reduced chilling injury. The efficiency of IVM/IVF technique in the porcine species is relatively lower than that obtained in other species such as ruminants. Two experiments were designed to investigate the effects of in-vitro fertilization of porcine oocytes matures using different OPS protocol for chilling and warming of vitrification. Porcine oocytes from ovaries collected at abattoir were matured for 44 hours in TCM199 Earle's salt supplemental with pyruvate, pff, L-cysteine, hormones and gentamycin. Oocytes were denuded and fertilized with frozen boar semen by common method. Porcine embryos produced routinely by in-vitro culture system of NCSU23 medium. The vitrification and the warming were conducted by OPS method with the glass micropipette instead of straw vessels and modified the protocol of G.Vajta(1999). In Exp 1, Chilling/Warming:Holding Medium(HM)+EG+DMSO/HM +sucrose Medium(SM) at 39$^{\circ}C$ warm stage. In Exp 2, : PBS+CS+EG+Ficoll+ Trehalose/PBS+Trehalose at 25$^{\circ}C$ stage. Filling, freezing, packing, thawing out and further culturing were performed to follow the basic protocol of G Vajta. During IVM-lVC and post-warming, fertilization parameter and developmental potential were compared to and statistically analysed. It was not significantly different from Exp 1 and Exp 2 but 25$^{\circ}C$ of stage was slightly higher on the morula/blastocyst forming rate and better atmosphere for worker than that at 39$^{\circ}C$ stage.

• 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.