• KOREAN JOURNAL OF CROP SCIENCE
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• 제37권2호
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• pp.134-140
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• 1992

Carrot(Daucus carota L.) seeds (CV Danver 126) were primed and then separated by density differences to improve both the percentage and time of radicle emergence. Seeds for priming were soaked in aerated distilled water for 2 days (water imbibed), 25% solution of polyethylene glycol(PEG) 6000 for 6 or 10 days, salt solution of 0.2M KNO$_3$＋0.1M $K_2$HPO$_4$ for 6 or 10 days, or mixed with Agro-Lig with 90% moisture content for 6 days and 70% moisture content for 6 or 10 days (SMP) at 2$0^{\circ}C$, respectively. The greatest embryo growth and the highest radicle emergence were observed from the seeds treated SMP with 90% moisture content for 6 days among the primed treatments. After the SMP treatment, the seeds were separated into density classes with a float-sink procedure using aqueous solution of Maltrin 600 with 0.02/cm$^3$ density increments. The lower density classes of the carrot seeds, the more embryo growth, the higher and the faster rates of radicle emergence were exhibited in order from 1.06 to 1.14 density classes of the carrot seeds treated SMP.

• Journal of Life Science
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• 제12권1호
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• pp.19-21
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• 2002

The pathway of embryos formed anther culture in herbaceous peony was influenced by addition of 2,4-D. MS medium with 2,4-Dichlorophenoxy acetic acid (2,4-D) alone did not arise direct embryogenesis, but was proliferate callus. Embryos through calli were produced on medium containing 0.2 mg/1 zeatin or without growth regulators. Direct embryogenesis was obtained from MS basal medium. However, after the anthers were cultured on medium with 0.1 mg/1 2,4-D, 3 g/1 AC, 30 g/1 sucrose, 2 g/1 gelrite for 40 days. Its efficiency (32.3 %) was markedly improved when anthers cultured on medium without 2,4-D. Embryo morphology was also affected by the 2,4-D used in medium. The induction of normal embryos with two cotyledons was higher in the embryos formed through direct embryogenesis than those formed callus. The embryos formed from calli were mainly showed abnormal embryo with one, three, four cotyledons or hors and bowling pin type.

• Journal of Embryo Transfer
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• 제27권3호
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• pp.171-174
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• 2012

Various small molecules can be used to control major signaling pathways to enhance stemness and inhibit differentiation in murine embryonic stem cell (mESC) culture. Small molecules inhibiting the fibroblast growth factor (FGF)/ERK pathway can preserve pluripotent cells from stimulation of differentiation. In this study, we aimed to evaluate the effect of pluripotin (SC-1), an inhibitor of the FGF/ERK pathway, on the colony formation of outgrowing presumptive mESCs. After plating the zona pellucida-free blastocyst on the feeder layer, attached cell clumps was cultured with SC-1 until the endpoint of the experiment at passage 10. In this experiment, when the number of colonies was counted at passage 3, SC-1-treated group showed 3.4 fold more mESC colonies when compared with control group. However, after passage 4, there was no stimulating effect of SC-1 on the colony formation. In conclusion, SC-1 treatment can be used to promote mESC generation by increasing the number of early mESC colonies.

• Journal of Embryo Transfer
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• 제26권2호
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• pp.111-115
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• 2011

In general, zona pellucida (ZP) of the blastocyst has to be removed first, then either isolated the inner cell mass (ICM) or ZP-removed whole blastocyst, which is then cultured on the feeder layer to induce ICM outgrowth for the generation of embryonic stem cells (ESC). However, it is unclear whether ICM isolation before seeding on feeder layer is beneficial or not because the interaction between ICM and trophoblasts may affect cellular growth and/or pluripotency during the culture on the feeder. In the present study, two ZP removal methods (mechanically by splitting with a 28-gauge needle versus chemically by the treatment of acid-Tyrode's solution) and two ICM isolation methods (ZP-free whole blastocyst seeding versus mechanical isolation of ICM) were evaluated for the efficient isolation and culture of putative parthenogenetic bovine ESC. The number of maintained outgrown colonies was counted in each experimental group. As the result, mechanical removal of ZP with a needle and followed by whole ZP-free blastocyst seeding on feeder cells tended to attach more on the feeder layer and resulted in more outgrown colonies with its simple and less time-costing benefits. Currently we are generating ESC lines in HanWoo cattle by using this method for initial outgrowth of the parthenogenetic bovine blastocysts.

• Asian-Australasian Journal of Animal Sciences
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• 제12권6호
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• pp.862-867
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• 1999

The present study was undertaken to determine the effect of administration of exogenous progesterone early in gestation on uterine levels of IGF-I mRNA and on embryo survival at day 25 of gestation in the pig. Forty-one prepubertal gilts were induced into oestrus with PG600 and artificially inseminated at their subsequent naturally occurring oestrus. Gilts were then randomly assigned to one of three groups. Gilts in the two treatment groups were injected intramuscularly with 50 mg of progesterone either from day 2 to 14 (N=14) or from day 4 to 14 (N=15) after breeding while those in the control group (N=12) were given corn oil (0.5 ml) from day 2 to 14. Between days 25 and 28 of gestation, gilts were slaughtered and reproductive tracts were recovered. Endometrial tissue (1 g) was collected and analysed for IGF-I mRNA levels using a reverse transcription-polymerase chain reaction, Progesterone treatment, starting either on day 2 or 4 after breeding, neither significantly increased embryo survival rate by day 25 of gestation nor altered IGF-I mRNA levels in uterine tissue. However, across all samples, the IGF-I mRNA level in the uterus was highly correlated with embryo survival rate (r=0.8193, p<0.01), supporting the involvement of IGF-I in the regulation of porcine embryo development.

• Proceedings of the Plant Resources Society of Korea Conference
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.60-60
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• 2018

This study was carried out to develop the seed propagation method of Dicenta spectabilis (L.) Lem. which is an ornamental plant native to Korea. In the previous studies, it was found that the seeds of D. spectabilis were morphophysiologically dormant (MPD), and high and low temperature of stratification were continuously required for the embryo growth and germination of the seeds. Especially, it was most effective to store for 1 month at $20^{\circ}C$ and then to transfer to $4^{\circ}C$. The treatment of $GA_3$ was carried out to promote embryo development and germination. The seeds were submerged in 100, 200 or $500mg{\cdot}L^{-1}$ $GA_3$ for 72 hours and then stored at various conditions as follow. The temperature conditions disposed of this experiment were 1 month at 10, 15, 20, and $25^{\circ}C$ or 2, 4, 8, and 12 months at $4^{\circ}C$, respectively. As a result, the length of embryo and germination rate of the seeds were the best when stored at $4^{\circ}C$ for 8 months after $500mg{\cdot}L^{-1}$ $GA_3$ treatment. Besides, when the seeds stored at $4^{\circ}C$, significant differences in embryo length and germination rate were shown with $GA_3$ concentration and storage period. It was also proved that high-concentration of $GA_3$ could replace the high temperature and could promote germination. Consequentially, the D. spectabilis seeds were classified into intermediate simple levels among MPD types.

• Toxicological Research
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• 제11권1호
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• pp.31-36
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• 1995

Cyclophosphamide (CP) must be enzymatically activated by cytochrome P450(CYP)-linked mixed-function oxidation pathway to be either mutagenic or teratogenic. Influences of alterations in hepatic mixed-function oxidase acitivity and glutathione (GSH) content on the embryotoxicity of CP were studied in rat whole embryo culture system. The embryotoxicity of CP was compared using rat S-9 fraction (S-9) pretreated with chemicals inducing different CYP isozymes, acetone (ACE), Aroclor 1254 (ARO), $\beta$-naphthoflavone (NAF) and phenobarbital (PHE). When 10.5 day embryos were cultured in the immediately centrifuged rat serum for 48 hrs using general gas char{ging schedule, CP$(40{\mu}g/ml)$ with S-9 induced by either NAF or PHE increased the incidence of realformations and significantly decreased embryonic growth compared with the non-induced S-9 group. ACE or ARO induced S-9 group showed no significant difference in embryonic growth. These data suggest that PB and/or NAF inducible CYP isoenzymes are mainly involved in the activation of CP. To examine the effect of GSH on the embryotoxicity of CP, 10.5 day embryos were exposed to CP and S-9 after preincubation with 10 mM of GSH for 3 hrs. In the GSH pretreated group the growth of embryos increased significantly compared with that of the untreated group, suggesting that GSH may protect embryos in culture from some toxic effects of CP.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.94-100
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• 2020

The inverse correlation between maternal age and pregnancy rate represents a major challenge for reproductive endocrinology. The high embryo ploidy error rate in failed in vitro fertilization (IVF) cycles reflects genetic misfires accumulated by older oocytes over time. Despite the application of different follicular recruitment protocols during IVF, gonadotropin modifications are generally futile in addressing such damage. Even when additional oocytes are retrieved, quality is frequently poor. Older oocytes with serious cytoplasmic and/or chromosomal errors are often harvested from poorly perfused follicles, and ovarian vascularity and follicular oxygenation impact embryonic chromosomal competency. Because stimulation regimens exert their effects briefly and immediately before ovulation, gonadotropins alone are an ineffective antidote to long-term hypoxic pathology. In contrast, the tissue repair properties (and particularly the angiogenic effects) of platelet-rich plasma (PRP) are well known, with applications in other clinical contexts. Injection of conventional PRP and/or its components (e.g., isolated platelet-derived growth factors as a cell-free substrate) into ovarian tissue prior to IVF has been reported to improve reproductive outcomes. Any derivative neovascularity may modulate oocyte competence by increasing cellular oxygenation and/or lowering concentrations of intraovarian reactive oxygen species. We propose a mechanism to support intrastromal angiogenesis, improved follicular perfusion, and, crucially, embryo ploidy rescue. This last effect may be explained by mRNA upregulation coordinated by PRP-associated molecular signaling, as in other tissue systems. Additionally, we outline an intraovarian injection technique for platelet-derived growth factors and present this method to help minimize reliance on donor oocytes and conventional hormone replacement therapy.

• Journal of Plant Biotechnology
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• 제42권4호
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• pp.376-379
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• 2015

This study was conducted to evaluate the effects of different types and concentrations of organic nitrogen sources (${\small{L}}$-Glutamine and casein hydrolysate, CH) and plant growth regulators (auxins and cytokinins) on embryogenic tissue proliferation and somatic embryo production in L. kaempferi. Overall, the highest tissue fresh weight was obtained at either 2 or 4 weeks in culture when 1,000 mg/L ${\small{L}}$-Glutamine was added to the culture medium, which showed similar results with other treatments. In experiments with different types and concentrations of plant growth regulators on somatic embryo production, the highest production (426.3/90 mg tissue) was found when 0.2 mg/L IBA was added; however, no somatic embryos were induced following treatment with 0.2 mg/L BA or Kinetin. The effect of various concentrations of IBA on somatic embryo production was also tested. The best result (303/90 mg tissue) was obtained when plants were treated with 0.2 mg/L IBA; 1.0 mg/L IBA was also effective (281/90 mg tissue). The lowest result (109.3/90 mg tissue) was obtained with 5.0 mg/L IBA.

• Korean Journal of Cleft Lip And Palate
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• 제4권1호
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• pp.13-26
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• 2001

Cleft lip and/or palate is the congenital orofacial malformation most commonly occurred in humans, The disease is multifactorial and is probably caused by genetic and/or environmental factors, So, there are many problems in research concerning etiology and in treatment of the disease, Even the most practiced and sophisticated methods of surgical repair are necessarily followed by scar contraction and fibrosis, which result in skeletal defects, dental abnormalities, cosmetic disfigurement, and speech impairment, As a result, Fetal surgery can be considered but practiced rarely when the deformity is not fatal to life, And treatment of cleft palate is performed in the form of medicine projection into uterus in animal experiments, Many studies show that growth factor and its receptor emerge from the developing palate; and the epidermal growth factor receptors have a important role in craniofacial development and in palatal fusion, The palatal morphogenesis of the avine is different from the mammal's; it takes the form of physiologic cleft palate, Recently, cleft palate fusion experiment was performed when the avine were in the period of palate formation through the exogenous TGF-β3 addition, and it showed that the exogenous TGF-β3 makes fusion of divided palate through certain process when cleft palate is occurred in palatal formation, In this study, I had the conformation of the fusion of cleft palate through the addition of TGF-β in case of chicken embryo, and observed the effect of TGF-β in EGF receptor distribution, And the following is the results of this study, 1. In case of the TGF-βl and β3 addition group, there was the decrease of EGFR(Epidermal Growth Factor Receptor) immunoreactivity in mesenchymal cells beneath the medial edge epithelium and also in epithelial mesenchymal interface which is between medial edge epithelium and nasal septum in 72 hours, 2, The immunoreactivity of the control group resembles that of normal chicken embryo palate in development, 3. In the view through fluorescence confocal microscopy, there was confluence in TGF-β3 addition group, This shows that the confluence induced by exogenous TGF-β3 is related to EGFR expression in palate of chicken embryo, which is a physiologic cleft palate model.