• 제목/요약/키워드: In-vitro culture

검색결과 2,898건 처리시간 0.031초

The Effects of Antioxidants on the Culture of Mouse Preantral Follicles In Vitro

  • Kim, Dong-Hoon;Kim, Dong-Kyo;Yang, Byoung-Chul;Park, Jin-Ki
    • Reproductive and Developmental Biology
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    • 제37권4호
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    • pp.193-197
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    • 2013
  • In order to investigate the effects of antioxidants on the culture of mouse preantral follicles in vitro, we examined the effects of taurine, glutathione and catalase on their growth and maturation. Addition of taurine was not effective on the survival of preantral follicles. However, metaphase II rates of oocytes within preantral follicles were significantly higher in 1 mM treated group than in control and 10 mM treated group (p<0.05). Glutathione did not improved the rates of survival and metaphase II oocytes. However, metaphase II rates of oocytes progressively decreased with increasing glutathione concentration. Catalase also showed that the rates of survival and metaphase II oocytes progressively decreased with increasing concentration. Especially, all of preantral follicles cultured in medium containing 100 IU/ml catalase were degenerated. These results suggest that low concentraion of taurine, as an antioxidant, have positive effect on the culture of mouse preantral follicles in vitro.

Optimization of in Vitro Cultivation of Inonotus Obliquus

  • Cho, Nam-Seok;Shin, Yu-Soo
    • Journal of the Korean Wood Science and Technology
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    • 제33권5호통권133호
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    • pp.92-98
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    • 2005
  • This study was performed to get the basic information concerned to the optimum culture condition of Inonotus obliquus. Several solid media, PDA, MEA and Czapek-Dox, and three liquid media were adopted for the in vitro cultivation. Some main features of the fungal morphological characteristics under cultivation conditions were observed and described. Preliminary results showed that appearance of the mycelial mat, hyphal size and substrate pigmentation differed according to the media. The PDA medium was the most favorable substrate for the growth on solid culture, followed by MEA and Czapek-Dox media. Concerned to the addition of amino acids, 5 amino acids, such as alanine, alginine, isoleucine, leucine and threonine, enhanced to the mycelial growth. Isoleucine was shown the best fungal growth. An important morphological hyphal structure for the fungus, the setae, was found in abundance and diverse its shape and size. In liquid culture, fresh potato broth was the best growth stimulant of the fungus, followed by Malt extract and potato broth. Addition of yeast extract to the liquid media had improved the biomass, but not laccase production.

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

  • Choi, Kwang-Hwan;Yoon, Ji Won;Kim, Minsu;Jeong, Jinsol;Ryu, Minkyung;Park, Sungkwon;Jo, Cheorun;Lee, Chang-Kyu
    • 한국축산식품학회지
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    • 제40권4호
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    • pp.659-667
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    • 2020
  • Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

체외생산 소 배반포의 이식에 의한 한우 쌍태 생산 (Production of HanWoo(Koran Native Cattle) Twin Calves by Transfer of Bovine Blastocysts Produced In Vitro)

  • 윤종택;이호준;노상호;정연길;손동수;김일화;류일선;김창근;정영채
    • 한국수정란이식학회지
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    • 제14권3호
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    • pp.171-176
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    • 1999
  • This study was carried out to investigate the effect of co-culture system(bovine oviduct epithelial cells; BOEC) and defined culture system (modified TALP ; mTALP) on the conception of embryos transferred, and pregnancy and twin birth rates after transfer of fresh or frozen-thawed bovine blastocysts produced in vitro were also evaluated. Oocytes from the slaughterhouse ovaries were matured and fertilized using general protocol. The results obtained were as the following. The pregnancy rate after transfer was higher in co-culture group than in mTALP group, but was not signficantly different, and there is no difference between fresh embryo group and frozen-thawed embryo group in conception rate. The conception rate was not different whether 3∼4 blastocysts or 2 blatocysts transferred into a recipient, but the production rate of twin calves was significantly higher (p<0.05) when 3∼4 embryos transferred. The average birth weight of twin calves(24.38kg) was numerically, but not significantly lighter than that of single calves(26.68kg).

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생쥐 체외수정 정도관리의 유용성에 관한 실험적 연구 (Mouse Embryo Culture as Quality Control for Human In Vitro Fertilization)

  • 임영경;박현정;이여일
    • Clinical and Experimental Reproductive Medicine
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    • 제18권1호
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    • pp.49-53
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    • 1991
  • The development of 2-cell mouse embryos to the blastocyst stage in vitro has been used as a quality control for the media empolyed for human in vitro fertilization. There was a comparison between the quality control data of the culture medium as ascertained by 2-cell mouse embryos development and sperm motility and the data from fertilization and cleavage of human oocytes. However, there was no obvious association between fertilization and cleavage of human oocytes and the quality of the medium ascertained by mouse embryo development and sperm motility.

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체외수정 및 미세조작에 의한 가축배의 생산가 효율적 이용에 관한 연구 I. 체외성숙.체외수정된 토끼 및 소 배의 이식과 동결 (Studies on Production and Efficient Utilization of Livestock Embryos by In Vitro Fertilization and Miceomanipulation I. Transfer and Freezing of In Vitro Fertilized Rabbit and Bovine Oocytes Matured In Vitro)

  • 김창근;정영채;이근상;김희석;정진태;윤종택;최미호;이장희;김광식
    • 한국가축번식학회지
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    • 제17권1호
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    • pp.57-68
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    • 1993
  • This stduy was carried out to find a reliable method for the production of in vitro fertilized embryos having more excellent development capacity and freezability in the rabbit and cattle. The greatest number of rabbit oocytes was recovered 6hrs after HCG injection(P<0.05). The maturation rate in vitro was slightly higher in the oocytes(6-h-oocytes) from 6h than those (8-h-oocytes)from 8 hrs after HCG injection and the beneficial effect of FSH during oocyte maturation was significantly great in the oocytes from large follicles. The cleavage rate into 2-to-6-cell stage was not differ between the 6-h-oocytes and 8h-oocytes, but the cleavage of these oocytes was greatly promoted by FSH addition to maturation medium and the cleavge of 8-h-oocytes matured without FSH was significantly low. The embryo development into 16-cell to morula was not promoted by the co-culture with rabbit oviduct epithelial cells. The freezability by embryo stages was ovidusly high at 4-cell and morula stage in 6-h-oocytes and the viability of 16-cell embryos from 8h-oocytes was similar to that of morula stage. The implantation sites after surgical tranfer of fresh rabbit embryos were not implanted. In bovine experiment, the in vitro development into 16-cell and morula after in vitro maturation and fertilization in the follicular oocytes was slightly improved by the co-culture with granulosa cells compared to that with oviduct epithelial cells and the frozen-thawed viability rate of these embryos ranged from 14 to 40%. The excellent fresh embryos were transferred nonsurgically to 6 recipients, but were not pregnant.

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돼지 난포란으로부터 배반포의 체외생산에 있어서 체외성숙시 기초배양액에 ESCM과 ESM의 첨가효과 (Effect of Addition of ESCM and ESM during In Vitro Maturation on In Vitro Development of Porcine Follicular Oocytes)

  • 김석기;박흠대
    • 한국동물생명공학회지
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    • 제34권3호
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    • pp.205-211
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    • 2019
  • In this study, we investigated the possibility of using mouse embryonic stem cell conditioned medium (ESCM) and embryonic stem cell medium (ESM) for in vitro maturation in the efficient in vitro production of blastocysts from porcine follicular oocyte. Depending on the concentration of supplement of ESCM added to the NCSU-23 solution did not affect 2-cell development rates and blastocysts development. However, in particular, the survival rate (10 days of culture) of blastocyst was significantly higher than that of the control group as the additive concentration (30%) increased (p < 0.05). The survival rate of blastocysts showed a similar tendency even with addition of ESM (30%) alone. On the other hand, the duration of the addition of these additives during IVM (0-44 h) was that the IVM I period (0-22 h) were more effective than the IVM II period (22-44 h). Thus, the effect of these additives is probably due to the combination of the various physiologically active substances of ESCM or the appropriate amino acids and vitamins of ESM. In particular, these additives were more effective during the first half (IVM I) of in vitro maturation. In summary, optimization of ESCM or ESM supplementation may improve in vitro maturation of porcine oocyte and affect developmental competency. Therefore, if more efficient methods of adding ESCM or ESM to basal culture medium can be developed during in vitro maturation of porcine follicle oocytes, high quality blastocysts will be developed from low porcine follicular oocyte compared to other domestic animals.

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
    • 한국수정란이식학회지
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    • 제27권4호
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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.