• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.399-407
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• 2001

The microinjection of retroviral vectors into the perivitelline spaces of MII-stage oocytes increased production of transgenic bovine embryos. However, oocytes have various sizes of perivitelline space, and there is the tendency that the oocyte membranes are damageable by micropipettes during the injection of retroviral vectors into perivitelline spaces or oocytes. Thus, it was not always possible to stably inject retroviral vector into perivitelline spaces of oocytes. Here we used sucrose to minimize the damage of the oocyte membrane. When the oocytes were suspended in 0.5% sucrose, poor quality oocytes showed rough cytoplasmic membranes, while good quality oocytes maintained smooth membranes. However, when the tatters were subjected to in vitro fertilization, no significant differences were observed in cleavage rates (82% of control Vs. 84% of sucrose treated oocytes). The Same trends were obtained from the oocytes fertilized after microinjection of LN$\beta$-EGFP and LNC-hGH genes into the perivitelline spares. The rates of cleavage and blastocyst from microinjection of LN$\beta$-EGFP genes were 81 and 25%, and from microinjection of LNC-hGM genes were 53 and 30%, respectively. The result indicated that microinjected oocytes could develop to the blastocyst stages after in vitro fertilization with no significant difference from control group. Moreover, the integration of hGH-gene (by PCR analysis) was detected in 52% of infected cleaved embryos and the expression of EGFP-gene (under a fluoresrence microscope) was also observed in 34% of infected blastocyst. These results indicated that 0.5% sucrose treatment could be an efficient method not only to select good quality embryos but also to inject retroviral vectors into perivitelline spares without any harm and hence improving developmental rates.

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.173-177
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• 1991

In vitro survival of the mouse morulae frozen by vitrification method(Kasai et al., 1990) was investigated in the present study. The embryos were plunged into LN2 directly after exposure to the vitrification solutions(EFS, GFS and DFS). The results were obtained as follows. The viability of morulae after freezing and thawing was high in EFS(96.7∼100.0%) and GFS vitrification solution(93.3∼96.7%), and the lowest in DFS vitrification solution(0.00∼0.03%).

• Macromolecular Research
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• v.11 no.1
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• pp.42-46
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• 2003

Cell attachment and proliferation on the polymer films of triblock copolymer(ester-ether)s comprising po1y (L-1actide) (PLLA) and poly (oxyethylene-co-oxypropylene)(PN) were investigated using 3T3 fibroblasts. It was found that on the tissue culture polystyrene(TCPS) and the PLLA control film the cells could spread well while on the copolymer films the cells showed a rounded morphology without spreading and proliferated weakly. Especially, little cells proliferated on the films of copolymer having a LN composition of 20 wt%. While the water absorption of the copolymer films increased with increasing PN content, the contact angle against water of copolymer films immersed in aqueous medium was almost identical, being slightly lower than that of the PLLA film. These properties were compatible with the results of cell attachment. The in vitro hydrolysis of the films of triblock and multiblock type copolymers was faster with increasing PN content. The increased hydrolyzability, the flexibility and the decreased cell attachment suggested that these copolymers may have high potential as biodegradable materials for medical use.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.19
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• pp.8107-8113
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• 2014

The aim of this study was to investigate the effects of olanzapine on growth inhibition as well as autophagy in glioma cells in vitro and in vivo. The proliferation of both LN229 and T98 glioma cells, measured by MTT assay, was suppressed in a concentration-dependent and time-dependent manner. Moreover, apoptosis of both cells was significantly increased with the treatment of olanzapine as evidenced by increased Bcl-2 expression, Hoechst 33258 staining and annexinV-FITC/PI staining. Olanzapine treatment also enhanced activation of autophagy with increased expression of LC3-II, expression of protein p62, a substrate of autophagy, being decreased. The growth inhibition by olanzapine in both glioma cell lines could be blocked by co-treatment with 3-MA, an autophagy inhibitor. Furthermore, olanzapine effectively blocked the growth of subcutaneous xenografts of LN229 glioma cells in vivo. The increased level of protein LC3-II and decreased level of p62 followed by a decreased level of Bcl-2, suggesting that autophagy may contribute to apoptosis. In addition, reduced proliferation of glioma cells was shown by a decrease of Ki-67 staining and increased caspase-3 staining indicative of apoptosis in mouse xenografts. These results indicated that olanzapine inhibited the growth of glioma cells accompanied by induction of autophagy and apoptosis both in vitro and in vivo. Olanzapine-induced autophagy plays a tumor-suppressing role in glioma cells.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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• pp.81-85
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• 1990

We cryopreserved mouse morulae by a simple ultra-rapid method of freezing embryos directly in $LN_2$ after holding 2min in a $LN_2$ vapor, and thawed them in $37^{\circ}C$ water bath. The time requirements for permeation and dehydration by 2.0 M glycerol and 0.2 M sucrose before freezing were studied. When the embryos were equilibrated for 10 min, the optimun post-thaw survival was obtained. Embryos those developed normally to blastocyst after in vitro culture for over 24hrs were regarded as survival ones. Two experiments to assess post-thaw survival following predehydration in various mixtures of glycerol and sucrose were also accomplished. When sucrose was held constant (0.2 M) and glycerol concentration varied (1.5-3.5 M), post-thaw survival was best (78.0%) in 3.0 M glycerol. When glycerol was held constant (3.0M) and sucrose concentration varied (0.0-1.0M), optimun post-thaw survival (78.0%) was found in 0.2 M sucrose.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.62-62
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• 2001

Recently, production of transgenic animal by nuclear transfer has been known as a useful method. The production of cloned offspring derived from nuclear transfer depends upon a variety of factors such as species, donor cells type and cell cycle, and source of recipient ova. Therefore, we attempted a different transgenic methods using follicular granulosa cells (GCs). In general, ovulated GCs undergoes lutenization and transformation in vitro which might defective effects on developmental potential. In order to avoid the GCs transformation in vitro culture system, we introduced a direct injection of retrovirus into the follicles and then collected them mechanically from ovaries of 6-8 week-old ICR mice. Retrovirus vector constructed with pLN $\beta$ EGFP was injected into the follicles. The follicles are cultured in $\alpha$ -MEM supplemented with human FSH, LH and ITS in Costar Transwell dish for 4 days. Survival rate of virus injected follicles was 52.1% (12/23) and expression rate of EGPP gene was 33.3% (4/12). In this study, we found GCs performed transgenesis in our culture system. In addition, the GCs in follicle may be developed in vivo like environment rather than in vitro environment. Thus, the use of GCs as donor cells may be useful in the nuclear transfer for cloning of genetic modification. Therefore, these results suggest that follicular GCs can be transfected by viral vector during folliculogenesis in vitro.

• Journal of Korean Society of Forest Science
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• v.95 no.5
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• pp.585-590
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• 2006

In vitro-grown axillary shot-tip meristems of Lycium chinense Mill. from cold-acclimated plant were successfully cryopreserved using a vitrification technique. After loading for 15 minutes with a mixture of 2.0 M glycerol and 0.4 M sucrose ($20^{\circ}C$), small segments (1-2 mm, 3-4 mm, and 5-6 mm) were cut from axilary buds and exposed to the cryoprotectant solution containing 30% glycerol, 15% ethylen glycol,15% dimethyl sulfoxide (DMSO), and 0.4 M sucrose at $0^{\circ}C$ for 30-120 minutes prior to direct plunge into liquid nitrogen (LN). After rapid thawing ($40^{\circ}C$), the segments were washed with MS medium containing 1.2 M sucrose for 0-35 minutes, and then transferred onto recovery-growth medium. The highest survival rate (about 90%) was obtained with cold-hardening treatment, and cryopreserved explants were successfully recovered to plantlets. No abnormal morphological changes were observed with the recovered plants after cryopreservation.

• Archives of Pharmacal Research
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• v.17 no.3
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• pp.150-153
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• 1994

Ethanolic extract of Daucus carota (DC) at the dose of 10-100 mg/kg caused a dose-dependent fall in systolic and diastolic arterial blood pressure in nomotensive anesthetized rats. These effects were not blocked by atropine (1 mg/kg) and pretratment with DC did not alter the pressor response to norepinephrine indicating that cardiovascular effects of DC are independent of cholinergic or adrenergic recptors involvement. In spontaneously beating guinea-pig paired atria, DC induced a concentration-dependent (03-5 mg/ml) decrease in force and rate of atrial contractions. In rabbit thoracic aorta, DC caused inhibition of $K^+$-induced contractions at similar concentrations. These results suggest that the extract may exhibit $Ca^{2+}$ channel blocking-like direct relaxant action on cardiac and smooth muscle preperations and this action may be responsible for its hypotensive effect observed in the in vivo studies.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.225.1-225.1
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• 2003

Purpose. Paclitaxel has demonstrated significant activity in clinical trials against a wide variety of tumors. The clinical application of Taxol$\^$\ulcorner/, a commercial product of solubilized paclitaxel with co solvents of ethanol and Cremophor, however, has been limited largely by hypersensitivity of the excipient. The aim of this study was to formulate paclitaxel-loaded lipid nanodispersions (Px-LN) for i.v. administration without toxic excipients, and to evaluate in vitro characteristics and in vivo pharmacokinetic behaviors. (omitted)

• Korean Journal of Plant Resources
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• v.29 no.6
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• pp.635-641
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• 2016

Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulation-vitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at $25^{\circ}C$. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.