• Title/Summary/Keyword: in vitro condition

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Conservation of An Endangered Species of Lilium cernum Komarvo. through in vitro Mass-Propagation (멸종위기 종 솔나리의 기내 대량생산을 통한 보존 연구)

  • Lee, Song-Hee;Lee, Su-Gwang;Kang, Ho-Duck
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.1
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    • pp.9-15
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    • 2011
  • This study was conducted to establish the optimal condition for conservation of genetic resources and micropropagation of Lilium cernum. Induction of bulbet of L. cernum was highly effective (9.2 bulb/explant) on 1/2 SH (Schenk and Hildebrandt) medium supplemented with 1.0 mg/L TDZ (Thidiazuron) and 0.1 mg/L NAA (Naphthaleneacetic acid). The treatment of 0.1 mg/L NAA increased root development (6.4 root/explant) under the in vitro condition. In addition, treatments of AC (Activated Charcoal) and ventilation were enhanced to develop number of shoots and to elongate length of leaf, bulb and root. Futhermore, the process of short-term soil acclimatization was promoted to strengthen the plantlets induced under the in vitro condition.

In vitro Follicular Growth and Ovulation of Mouse Preantral Follicles Cryopreserved by Vitrification (초자화동결된 생쥐 Preantral Follicle의 체외성장과 배란)

  • Park, Ji-Kwon;Paik, Won Young
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.91-99
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    • 2005
  • Objective: To define an appropriate vitrification condition of preantral follicle that yields high survival and to evaluate growth and ovulation rate of mouse follicles during in vitro culture after vitrification. Methods: Preantral follicles were isolated mechanically from mouse ovaries that were surgically recovered from mice aged 14 days. Retrieved preantral follicles were placed in EG (Ethylene Glycol) for 2, 5, 10 minutes and transferred to EFS-40 (40% EG, 18% Ficoll-70, 0.5 M sucrose) for 0.5, 1, 2 minutes. And then, preantral follicles were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing was carried out at room temperature. After defining the most appropriate vitrification condition that yields high survival, in vitro growth and ovulation rate of follicles were evaluated. Results: Appropriate vitrification condition that yield high survival rate ($83.2{\pm}2.1%$) of preantral follicle was EG for 5 minutes and EFS-40 for 0.5 minutes. In vitro survival rate of the vitrified preantral follicles were $85.5{\pm}0.5%$, $67.9{\pm}0.8%$ and $40.2{\pm}0.5%$ on day 2, 6 and 10. And in vitro growth of the vitrified preantral follicles were $107.1{\pm}16.1{\mu}m$, $117.1{\pm}18.4{\mu}m$, $178.4{\pm}45.6{\mu}m$ and $325.4{\pm}54.4{\mu}m$ on day 0, 2, 6 and 10. Although in vitro survival rate and growth of vitrified preantral follicles were lower than that of non-vitrified preantral follicles, the patterns of survival and growth were similar in vitrified and non-vitrified preantral follicles. The ovulation rate of antral follicles that was grown from vitrified preantral follicles was $32.6{\pm}1.2%$. Conclusion: Vitrified preantral follicles could be grown to antral sizes, and mature oocytes that can be used for IVF-ET programs were produced successfully. These data suggest that cryopreservation of preantral follicle by vitrification can be used for the preservation of the fertility.

Viability of In Vitro Fertilized Bovine Embryos Following In Vitro Culture and Embryo Transfer (소 체외수정란의 체외배양 및 이식후 생존성)

  • 정희태;유재원;박연수;양부근;김정익
    • Journal of Embryo Transfer
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    • v.9 no.3
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    • pp.221-227
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    • 1994
  • This study was conducted to examine the condition of in vitro culture system and the viability after embryo transfer of in vitro matured-in vitro fertilized (IVM-IVF) bovine embryos. The in vitro development to the blastocyst stage was enhanced by supplying bovine serum albumin(BSA) to co-culture medium with bovine oviduct epithelial tissue(BOET) compared with that in medium supplemented with fetal bovine serum(FBS) (41.2% vs. 26. 3%, P<0.05). After transfer of IVM-IVF blastocysts into the uterine horn of recipient females (Aberdeen Angus), one was pregnant to term and produced a head of male Korean native calf. These results confirm that the in vitro development of IVM-IVF bovine embryos is affected with different protein source in co-culture with BOET, and IVM-IVF embryos can develop to term after in vitro culture and embryo transfer.

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Study on Germination of Seed and Growth of Rhizome in Cymbidium goerinii in vitro (Cymbidium goeringii 종자의 in vitro 에서의 발아에 관한 기초적 연구)

  • EuiSooYoon
    • Korean Journal of Plant Resources
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    • v.2 no.1
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    • pp.235-241
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    • 1989
  • The study was conducted to determine the Ms orthogonaL modia and the concentration of plant growth regulator for seed matura-tion and growth of rhizome from Cymbidium goeringii Germination waswell in dark condition, but the growth of rhizome was better un-der dark than under light condition in MS orthoTonal . Sucrose con-centration( 3 %) gave better results than higher ones(6%), andthe use of NAA(0.1 PPm) effect significant difference of seed ge-rmination .But the growth of rhizome was best in medium Containingsucrose concentration(3%) Ippm NAA and 1 PPm BA.

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Studies on In Vitro Culture, Freezing and Transfer of Korean Native Cattle Embryos Fertilized In Vitro H. Factors Affecting on Survival Rate of Frozen-Thawed Korean Native Cattle Embryos Fertilized In Vitro (한우 체외수정란의 체외배양, 동결보존 및 이식에 관한 연구 II. 한우 체외수정란의 동결 및 융해 후 생존율에 영향을 미치는 요인)

  • 김일화;손동수;이호준;최선호;양병철;이광원;장인호
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.125-135
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    • 1996
  • The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

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A Clinical Report of an Infertile Patient with Adenomyosis Who Failed Repeatedly In Vitro Fertilization (체외수정에 반복적으로 실패한 자궁선근증을 동반한 난임환자 치험 1례)

  • Jung, Myung-Ju
    • The Journal of Korean Obstetrics and Gynecology
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    • v.30 no.4
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    • pp.203-212
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    • 2017
  • Objectives: To report the efficacy of traditional Korean medicine to an infertile patient who repeatedly failed in vitro fertilization. Methods: The patient was diagnosed with adenomyosis and failed in vitro fertilization 9 times. Her dysmenorrhea and physical symptoms were improved through traditional Korean medicine and she was pregnant with the 10th attempt of in vitro fertilization. She had bleeding during pregnancy due to adenomyosis and took herbal medicines to maintain stable condition. Results: During the treatment period, the uterine thickness due to adenomyosis was reduced and her dysmenorrhea was improved. She was pregnant by in vitro fertilization and gave birth to a healthy child by Caesarean section. Conclusions: This case report shows that traditional Korean medical treatments work to improve the success rate of in vitro fertilization.

Optimized Shoot Induction and Histological Study of in vitro Cultured Korean Soybean Cultivars

  • Kantayos, Vipada;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.32 no.3
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    • pp.237-243
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    • 2019
  • Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, 'Dawon', 'Pungsan', 'Daewon', 'Taekwang' and 'Chongdoo 1' by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, 'Dawon' and 'Pungsan'. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 mg/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in 'Dawon' cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

Effects of Strains of Saccharomyces cerevisiae and Incubation Conditions on the In vitro Degradability of Yeast and Roughage

  • Ando, S.;Nishiguchi, Y.;Hayasaka, K.;Yoshihara, Y.;Takahashi, J.;Iefuji, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.3
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    • pp.354-357
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    • 2005
  • The in vitro degradability of yeast and the effect of yeast on the in vitro degradability of forage may differ in terms of the specific yeast strains or their incubation conditions. Thus in experiment 1, two strains of sake yeast (strainK7 and strainK9) and one strain of bakers' yeast (KY5649) were incubated in an aerobic condition. In experiment 2, aerobically or anaero bically incubated K7 was used for investigating the in vitro degradability of yeast, the effect of yeast on the in vitro degradability of forage, and the degradability of yeast by pepsin and pronase treatment. The in vitrodegradability of bakers' yeast was significantly (p<0.05) higher than those of sake yeasts. The in vitro degradability of anaerobically incubated yeast was significantly (p<0.01) higher than that of aerobically incubated yeast. The degradability of bakers' yeast by pepsin treatment was significantly (p<0.01) higher than that of the sake yeasts. The degradability of bakers' yeast by pronase treatment was slightly higher than that of the two sake yeasts, while the degradability of anaerobically incubated yeast by both enzymes, respectively, was significantly (p<0.01) higher than that of aerobically incubated yeast. The degradability of forages was increased significantly (p<0.05) by the addition of yeasts. The degradability of roughage by sake yeast tended to be higher than that by the bakers' yeast. The degradability of roughage was significantly (p<0.05) higher by anaerobically incubated yeast than by aerobically incubated yeast. Given the above results, it seems that in vitro degradability of yeast and the magnitude of the increment of roughage degradation differ among the yeast strains and their incubation conditions.

Effects of Ovary Status and In Vitro Maturation Condition on the Developmental Competence of Canine Oocytes

  • Cho, Su-Jin;Kim, Dong-Hoon;Min, Chan-Sik;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.27 no.4
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    • pp.265-270
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    • 2012
  • In canine, oocytes are ovulated at the GV (germinal vesicle) stage and they have to fulfill maturation phase before reaching metaphase II stage. The efficiency of in vitro maturation is still very low. Therefore, the aim of this study was to investigate the effect of in vitro maturation on nuclear changes of immature canine oocytes recovered from different reproductive stages ovaries and different culture conditions. The oocytes were cultured in TCM-199 with supplement at 5% $CO_2$ and $38.5^{\circ}C$ for 72 h. The nuclear maturation of canine oocytes was evaluated with Hoechst 33342 stain under fluorescence microscope (Fig. 1). The results of this study detected differences in in vitro maturation rate between oocytes recovered from follicle status and non-follicle status ovaries. However, these differences were not significant as indicated in Table 1 and Fig. 2. In regard to the effect of culture condition with supplements, we did not found significant differences compared with control group (Table 2, Table 3). One of the reasons for this data could be the conditions that ovaries were exposed during slaughtering process or the long distant transportation of the ovaries. Although these data have not shown clearly significant differences results compared with control, furthermore the different reproductive status ovaries was beneficial for maturation of oocytes in vitro and can be a basic part of knowledge to improve in vitro maturation of canine oocytes.

Effects of Storage Duration, Medium and Viscin on in vitro seed Germination in Endangered Species, Loranthus tanakae (저장기간, 배지종류 및 viscin이 멸종위기종 꼬리겨우살이의 기내 종자발아에 미치는 영향)

  • Lee, Su-Gwang;Lee, Song-Hee;Kang, Ho-Duck
    • Journal of Korean Society of Forest Science
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    • v.99 no.4
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    • pp.618-624
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    • 2010
  • This study was conducted to establish the in vitro seed germination of Loranthus tanakae. A factorial experiment evaluated the effects of seed storage duration (0, 8, 16 weeks), media (MS, SH, White, WPM), presence of viscin, GA3, gelling agent and concentrations. Seed germinated after one week culture in in vitro condition and produced radicles. In vitro seed germination was optimal when the seeds removed viscin placed on SH medium (69%) and the addition of 0.35% gelrite (75%) in the same medium. As seed storage duration was expanded to 8 or 16 weeks, in vitro seed germination rate was reduced rapidly. Holdfasts were also produced at the side of radicles. The important factors to produce holdfast and haustorium was kind of media as an optimal condition in White medium without any supplements to be shown 98% and 8% respectively. Process of in vitro germination of Loranthus tanakae was followed to radicle elongation, holdfast development and then haustorium formation sequencially.