• Korean Journal of Pharmacognosy
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• v.49 no.1
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• pp.28-39
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• 2018

This study investigated the effect of snail extract on the growth parameters of old female rats (27 weeks). Rats were administered orally with snail extract at a dose of 100 mg/kg, 200 mg/kg, chondroitin sulfate 10 mg/kg and 0.9% saline (control) for 8 weeks. Bone mineral density (BMD) and serum concentrations of insulin-like growth factor 1 (IGF-1) and insulinlike growth factor-binding protein 3 (IGFBP-3) were significantly higher in rats exposed to snail extract for 8 weeks. MG-63 cells (human osteoblast-like cells) were treated with snail extract for 48 h. Their differentiation and proliferation was investigated with Western blot and morphological changes observed via immunofluorescence staining of ${\beta}-catenin$. Treatment with snail extract significantly increased the levels of growth factors including ${\beta}-catenin$ and IGF-1. The snail extract affected osteoblast formation. Morphological changes in MG-63 cells were observed via immunofluorescence staining. Treatment with snail extract increased the expression of ${\beta}-catenin$ in MG-63 cells. Results suggest that the treatment of MG-63 cells with snail extract increased the longitudinal growth and growth factor levels. Snail extract may be pharmacologically effective in osteogenic differentiation in vitro and represents a potential therapeutic agent for bone formation.

• Journal of Korean Society of Forest Science
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• v.95 no.2
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• pp.155-159
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• 2006

An efficient method for in vitro propagation of the medicinal plant Hovenia duleis, was established. Plantlets for micropropagation of H. dulcis were obtained from in vitro germinated seeds. The effectiveness of various levels of cytokinins (BAP, Kinetin and TDZ) on multiple shoot formation from stem nodes was tested. BAP (1.0 mg/L) treatment induced highest number of multiple shoots. The growth pattern of plantlet on various culture media was undertaken. The shoot elongation was optimal on 2MS basal medium without growth regulators. The in vitro rooting ability of H. dulcis shoots was examined with two-auxins IAA and IBA. The IAA (1.0 mg/L) treatments induced earliest rooting with maximum number of roots and root growth. Rooted shoots were transferred directly to small pots with artificial soil and such established plant exhibited a normal growth pattern similar to wild plantlet.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.2
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• pp.117-122
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• 2019

The objective of this study was to establish an in vitro culture system for ovarian preantral follicles of B6D2F1. First, we optimized the in vitro preantral-follicle culture by culture duration, follicle stimulating hormone (FSH) type, and activin A concentration. Duration of in vitro culture for 9, 11, and 13 days was sufficient for the normal development of preantral follicles to antral follicles. Formation of cumulus cell-oocyte complex (COC) was induced by treatment with human chorionic gonadotropin (hCG; 2.5 IU/mL) and epidermal growth factor (EGF; 5 ng/mL). In addition, metaphase II (MII) oocytes formed during this in vitro culture of preantral follicles. In vitro preantralfollicle culture for 9 days showed higher rates of growth and maturation, thus yielding a greater number of antral follicles, and there were significant differences (p < 0.05) in the number of MII oocytes (that formed from these preantral follicles via differentiation) between the 9-day culture and 11-day or 13-day culture. The follicles cultured for 9 days contained a tightly packed well-defined COC, whereas in follicles cultured for 11 days, the COC was not well defined (spreading was observed in the culture dish); the follicles cultured for 13 days disintegrated and released the oocyte. Second, we compared the growth of the preantral follicles in vitro in the presence of various FSH types. There were no significant differences in the growth and maturation rates and in differentiation into MII oocytes during in vitro culture between preantral follicles supplemented with FSH from Merck and those supplemented with FSH from Sigma. To increase the efficiency of MII oocyte formation, the preantral follicles were cultured at different activin A concentrations (0 to 200 ng/mL). The control follicles, which were not treated with activin A, showed the highest rate of differentiation into antral follicles and into MII oocytes among all the groups (0 to 200 ng/mL). Therefore, activin A (50 to 200 ng/mL) had a negative effect on oocyte maturation. Thus, in this study, we propose an in vitro system of preantral-follicle culture that can serve as a therapeutic strategy for fertility preservation of human oocytes for assisted reproductive medicine, for conservation of endangered species, and for creation of superior breeds.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.15 no.2
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• pp.53-79
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• 2002

To screen the effective materials for hair loss treatment, several natural extracts were tested using in vivo and in vitro test models. Firstly, all test materials were applicated onto the back skin of C57BL／6 mouse and then hair growth promoting effect was measured using hair growth index. As a result, Prunus mume, black bean, Brassica campestris subsp. black sesame and Rubi Fructus showed potent hair growth promoting effect, ranking as 1.5-2.0 of hair growth index. However, there were no plant extracts, which have remarkable potential of growth promotion of human hair dermal papilla cells cultured in vitro. In the experiments of 5${\alpha}$-reductase type Ⅱ inhibition assay, Prunus mume, Eriobotryae Folium showed effective potential to inhibit the activity of 5${\alpha}$-reductase type Ⅱ. To investigate the possible involvement of the effect of several plant extracts on the gene expression of growth factors in human hair dermal papilla cells, RT - PCR analyses were performed. However, there were no plant extracts, which have profound effect on the gene expression of several growth factors such as IGF-I, KGF, HGF and VEGF in the dermal papilla cells. Another tests for inhibition of microbial such as P. acne were also carried out to find whether these plant extracts have anti -microbial activities. Rubi Fructus showed anti -microbial effects on Propionibacterium acnes, which is believed as a pathogen of acne. Together, these results showed several plant extracts can be used for hair growth promotion.

• Journal of Bio-Environment Control
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• v.21 no.2
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• pp.134-139
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• 2012

Inhibition effects on spore germination and mycelia growth for gray mold (Botrytis cinerea) were investigated in vitro using environment-friendly agricultural materials as well as environment-friendly pesticides. The inhibition effect on mycelia growth of gray mold is the highest when the gray mold mycelia were treated with a pesticide (commercial name: Koreayeok, Jihabudea KM, Sootingtan, Sootingstar) that contains a mixture of Bacillus subtilis, resulting in 100% inhibition of the mycelia growth. Meanwhile, the range of less than 20% inhibition effects on the growth of gray mold mycelia was observed with other commercial agricultural materials. The significant inhibition effects on spore germination of gray mold fungus were shown in vitro with two water dispersible pesticides containing sulfur [BTB (97.7%) and SulfurStar (92.3%)], respectively. These in vitro results of inhibiting of the spore germination and mycelia growth together cannot found. It remains to be determined whether the selected environment-friendly agricultural materials in effective control of gray mold in vitro can be used to control gray mold in field.

• Mycobiology
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• v.30 no.2
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• pp.70-75
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• 2002

The mycelia were directly isolated from eight species of fungal basidiocarps, confirmed to the ectomycorrhiza in the roots from the fields(forestry); Suillus bovinus, Paxillus involutus, Lactarius hysginus, Russula fragilis, Lepista nuda, Lyophyllum shimeji, Tricholoma matsutake, and Russula integra. The mycelia were pure-cultured with several transferring in various agars, and inoculated to the roots of pine(Pinus densiflora) seedling by in vitro method. After ten months growth under artificially aseptic conditions, all pine seedlings inoculated were stimulated at the growth-height, whereas those not inoculated were nearly dead. Also, the ramifications of ectomycorrhizal pine roots formed in the synthetic in vitro systems and were various according to the different mycelia. Synthesis of ectomycorrhiza were clearly confirmed in ten months growth, but not distinguished at this moment. It was clearly proved that the mycelia isolated caused the ectomycorrhizae in the roots of pine seedlings.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.292-297
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• 1995

We developed an in vitro assay method for evaluating plant growth promotion and providing an evidence that the growth promotion is rendered by growth enhancing factors. The amendment of culture filtrates of Trichoderma harzianum T95 and Gliocladium virens G872 and G872B in Murashige and Skoog (MS) agar medium enhanced the cotyledon growth of cucumber in terms of fresh weight and primary leaf area of cucumber cotyledon cuttings, of which the treatment of G. virens G872B was the most effective. The mycelial culture filtrate of G872B was more effective in the growth promotion than its conidial germling filtrate. The addition of 1% sucrose in MS mineral medium with 0.1% culture filtrates of the antagonists (T95 and G872B) was optimum for enhancing the effect of the filtrates on the growth of cotyledon cuttings in vitro. When cucumber seeds treated with G872B, Pseudomonas putida Pf3 or the G872B-Pf3 mixture were planted in a greenhouse, the rate of seed germination, biomass of shoot and root, and yield of cucumber fruits were increased, especially by G872B or the G872B-Pf3 mixture. Correspondingly, cucumber fruit yields in early to middle-cycles of harvest were significantly greater in the plots of G872B than the control and Pf3-treated plots, and the final yield was highest in the plots of the G872B-Pf3 mixture applications.

• Journal of Plant Biotechnology
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• v.4 no.4
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• pp.151-154
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• 2002

Conditions for lily pollen growth in vitro and transformation were optimized. Active pollen tube development was achieved effectively in a medium containing 7% sucrose with pH adjusted to 5.7 at the temperature of 27$^{\circ}C$ for about 16-24 hours. Pollen growth was little impaired by the presence of kanamycin at concentration up to 100 mg/L. Pollen rains near the beginning of germination stage were more reliable for Agrobacterium-mediated GUS DNA transformation via vacuum infiltration lasted for 20-40 minutes. GUS DNA integration and its expression in fully developed pollen tubes could be confirmed by Southern blot hybridization, RT-PCR and histochemical staining.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.1
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• pp.35-39
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• 2019

This study aimed to recover the ovarian function through in vitro culture of preantral follicles from aged mice. First, we isolated the preantral follicles from ovaries of sixty-seven-week old B6D2F1 mice with decreased fecundity to know how many follicles were present in them, which was 6 preantral follicles including 2 primary, 2 early secondary and late secondary follicles from 8 aged mice. It was confirmed that a few follicles (~2) were present in aged mice through histological analysis compared to adult mice as control. The 9 days of in vitro culture of preantal follicles showed in vitro growth and induced maturation after treatment with hCG (2.5 IU/mL) and EGF (5 ng/mL). Cumulus cells in the cumulus-oocyte complexes (COCs) were removed using hyaluronidase and oocytes at the germinal vesicle (GV) and GV breakdown (GVBD) were obtained from preantral follicle culture of aged mice in vitro. In conclusion, these observations demonstrated that there still were a few preantral follicles in the ovaries of 67 week-old mice, which we were able to culture in vitro and oocytes were obtained from them. This study proposed an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation in humans for assisted reproductive medicine.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.119-126
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• 2009

An efficient in vitro plant regeneration system was developed through shoot proliferation from axillary buds of Tectona grandis (L.f), APNBV-1 (Andhra Pradesh North Badrachalam Venkatapuram-1) clone. Multiple shoots of high quality were produced in vitro from axillary bud explants. An average of 4.39 shoots/explant were obtained on Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) benzyl amino purine (BA), kinetin (KN), indole acetic acid (IAA), gibberillic acid ($GA_3$), growth adjuvants casein hydrolysate (CH), adenine sulphate (Ads) and antioxidants ascorbic acid, polyvinyl pyrrollidine (PVP). Eighty five percent of rooting was observed in ex vitro rooting media containing IBA and vermiculite. In ex vitro rooting, single shoots with 2 to 3 nodes were subjected to IBA of different concentrations at different periods of time intervals. Direct rooting in vermiculite at 500 ppm concentration of IBA resulted in 4.3 number of roots with 2 cm length. Minimum response of rooting and length of roots were recorded at 100 ppm concentration of IBA. Planlets were transferred to plastic bags for short acclimatization stage in green house where they survived at 95%.