• 제목/요약/키워드: in vitro preservation

검색결과 191건 처리시간 0.026초

Guidelines for Manufacturing and Application of Organoids: Skin

  • Seunghee Lee;Yeri Alice Rim;Juryun Kim;Su Hyon Lee;Hye Jung Park;Hyounwoo Kim;Sun-Ju Ahn;Ji Hyeon Ju
    • International Journal of Stem Cells
    • /
    • 제17권2호
    • /
    • pp.182-193
    • /
    • 2024
  • To address the limitations of animal testing, scientific research is increasingly focused on developing alternative testing methods. These alternative tests utilize cells or tissues derived from animals or humans for in vitro testing, as well as artificial tissues and organoids. In western countries, animal testing for cosmetics has been banned, leading to the adoption of artificial skin for toxicity evaluation, such as skin corrosion and irritation assessments. Standard guidelines for skin organoid technology becomes necessary to ensure consistent data and evaluation in replacing animal testing with in vitro methods. These guidelines encompass aspects such as cell sourcing, culture techniques, quality requirements and assessment, storage and preservation, and organoid-based assays.

In vitro에서 Thelohanellus kitauei 포자의 운명에 관한 지견보유 (Supplemental knowledge on survival of Thelohanellus kitauei spores in vitro)

  • 이재구
    • Parasites, Hosts and Diseases
    • /
    • 제32권1호
    • /
    • pp.57-60
    • /
    • 1994
  • Thelohanellus kitauei 포자의 운명을 조사하기 위하여 in vitro에서 극사탈출률을 경시적으로 조사하였다. 포자를 0.45% 및 0.9%식염수, 증류수 그리고 Tyrode액에 현탁시켜 $-70^{\circ}C$에 냉동하여 장기간 보존한 바 거의 모든 포자의 활성은 최초 냉동후 1.750일 째까지 초기와 거의 같았다. 0.45% 및 0.9%식염수 그리고 증류수에 현탁시켜 $5^{\circ}C$에 보존한 포자는 각각 1,628일, 1,614일 그리고 1,721일, 항생제를 첨가한 위의 현탁액에서는 각각 1,628일, 1.614일 그리고 1,714일에 모든 포자가 사멸하였다. 한편, Tyrode액에 현탁시킨 것은 모두 235일에 모든 포자가 사멸하였다.

  • PDF

Post-Thaw Cryosurvival of Bovine Embryos Produced In Vitro and In Vivo after Controlled Freezing

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Lee, Poong-Yeon;Son, Jun-Kyu;Kim, Jae-Bum;Kim, Sung-Jae;Kim, Hyun-Jong;Shin, Seung-Oh;Son, Dong-Soo
    • 한국수정란이식학회지
    • /
    • 제24권4호
    • /
    • pp.253-257
    • /
    • 2009
  • To enhance the embryo preservation technology and better application of embryo transfer technique to the field (dairy science or animal reproduction. etc.), we examined the viabilities of bovine embryos produced in vitro and in vivo after cryopreservation according to their developmental stage and thawing temperature. Bovine embryos from in vivo/vitro fertilization (Hanwoo) were examined at day 7, 8, and 9. Survival rates and total cell numbers of in vivo fertilized embryos were as follows: morulae 68.8% and $67\;{\pm}\;6.0$; blastocysts 80.5% and $120\;{\pm}\;10$; expanded blastocysts 77.4% and $138\;{\pm}\;9.7$, respectively. Rates of embryo development for blastocysts and expanded blastocysts after thawing were significantly higher than that of morula stage embryos (p<0.05). While survival rates of in vitro fertilized embryos according to developmental stage showed no significant difference among groups (morula 67.9%; blastocyst 74.3%; and expanded blastocyst 79.4%), total cell numbers were significantly lower than those of other groups (morula $64\;{\pm}\;5.9$; blastocyst $116\;{\pm}\;8.7$; and expanded blastocyst $135\;{\pm}\;9.1$) For the viability according to thawing temperature, survival rate was higher in $37^{\circ}C$.

Regeneration of Cryopreserved Pear Shoot Tips Grown in Vitro by Encapsulation-Dehydration

  • Yi, JungYoon;Lee, YoungYi;Lee, GiAn;Son, EunHo;Park, HongJae
    • 한국자원식물학회지
    • /
    • 제30권6호
    • /
    • pp.612-617
    • /
    • 2017
  • The preservation of pear germplasm, like that of other clonal germplasms, is difficult because it requires conservation of whole plants or their tissues. Among the currently available methods for long-term conservation of clonal germplasm, cryopreservation of shoot tips is the most reliable and cost- and space-effective option. Alginate-coated axillary shoot tips from in vitro-grown pear were conserved successfully in liquid nitrogen (LN) following dehydration. Shoot recovery from cryopreserved shoot tips was improved greatly after 8 weeks of cold acclimation, but recovery decreased slightly after then. The highest regeneration rate was observed when in vitro shoot tips were preincubated in MS (Murashige and Skoog) medium with 0.3 M sucrose for 48 h, and when alginate-coated shoot tips were precultured in MS medium with increasing sucrose concentrations (0.5 M and 0.7 M) for 8 and 16 h, respectively. When the encapsulated beads were dehydrated for up to 7 h [25% water content (fresh weight basis)] under laminar flow, the highest regeneration rate was observed in "BaeYun No. 3" (55.7%) and "Whanggeum" (43.3%) after warming from LN. This technique is useful as a practical procedure to cryopreserve plant material that is sensitive to freezing of the surrounding cryoprotectant medium. Therefore, this technique appears to be promising for the cryopreservation of shoot tips from in vitro-grown plantlets of pear germplasm.

효소 및 미생물 복합체를 사용한 인비트로 루왁 커피의 품질 특성 (Quality characteristics of in vitro luwak coffee produced using enzyme and microbial complexes)

  • 강혜미;오신영;강혜민;권중호;정용진
    • 한국식품저장유통학회지
    • /
    • 제30권2호
    • /
    • pp.287-299
    • /
    • 2023
  • 효소 및 미생물 복합제를 사용하여 인비트로 발효 루왁커피를 제조하여 non-fermented coffee beans(NFC)과 fermented coffee beans(FC)의 커피 품질을 비교하였다. 총유리아미노산 함량은 NFC가 254.01±4.89 mg/mL, FC가 264.15±16.80 mg/mL로 FC의 함량이 다소 높은 것으로 확인되었다. 이때 NFC는 glutamic acid, γ-aminon-butyric acid 등이 높았으나, FC는 lysine, leucine, valine 등 필수아미노산의 함량이 높았다. 커피 생두의 발효과정은 sucrose의 감소와 fructose 및 glucose의 유의적인 증가를 가져왔다. 커피 추출물의 색도는 NFC에 비해 FC 시료에서 높은 명도(L)와 적색도(a) 및 황색도(b)를 보였다. 카페인 함량은 NFC 1,130.22±1.55 ㎍/mL, FC 696.94±0.04 ㎍/mL로써 발효 후 약 38%의 카페인이 감소되었다. 폴리페놀 및 클로로겐산 함량은 NFC에서 각각 2.31±0.01 mg GAE/mL와 531.81±27.32 ㎍/mL, FC에서 각각 2.03±0.07 mg GAE/mL와 264.46±2.47 ㎍/mL로 발효에 따라 떫은맛 관련 성분의 함량이 유의적으로 감소됨을 알 수 있었다. 전자코 분석에서 NFC와 FC의 휘발성 향성분의 차이가 뚜렷함이 확인되었고, methylethyl formate, 2-methyl-1, 3-cyclopentadiene, 2-chloro-2-methylbutane, 2-methylbutanal 등의 휘발성 화합물이 발효 후 높은 강도로 감지되었다. 관능 평가에서는 NFC 추출물보다 FC 추출물의 aroma, body, aftertaste, overall에서 높은 관능 평점을 보여주었다(p<0.001). 이상의 결과에서 효소 및 미생물 복합제를 사용한 커피 생두의 발효는 유효 성분들의 변화를 일으켜 커피 원두의 로스팅 과정 중 Maillard 반응을 촉진함으로써, 향미가 증가되고 카페인 함량이 감소된 인비트로 루왁 커피의 제조 가능성을 시사하였다.

B16 흑색종에 대한 팽이버섯 추출물의 항암활성 비교 (Analysis of Anti-Tumor Activity of Flammulina velutipes Extract on B16 cells)

  • 이시림;남동윤;이현진;박철홍;허진철;김종국;이진만;이창윤;박희주;이상한
    • 한국식품저장유통학회지
    • /
    • 제16권4호
    • /
    • pp.599-603
    • /
    • 2009
  • 버섯은 예전부터 단백질, 당질, 비타민, 무기질 등 일반 영양소들이 다량 함유되어 있을 뿐 아니라 다양한 약리작용을 가지고 있기 때문에 예로부터 민식용 및 민간요법으로도 많이 이용되어 왔다(17). 또한 현재에는 건강식으로 다양하게 이용되고 있다. 본 연구는 여러 가지 버섯 중, 팽이버섯의 추출물을 이용하여 이의 항암 활성을 실시하였다. 팽이버섯(Flammulina velutipes)의 항암활성을 검증하기 위하여 in vitro 실험으로 wound healing 활성 실험을 하였으며, in vivo 실험으로 마우스를 이용한 항암모델 실험을 실시하였다. Wound healing 활성 실험 결과 팽이버섯 추출물의 농도를 0-100 ug/mL로 처리함에 따라 B16 세포의 운동성이 감소하는 것을 알 수 있었다. 이를 바탕으로 마우스를 이용하여 항암 모델에 적용한 결과 B16 세포로 인해 형성되는 colony가 팽이버섯 추출물을 처리한 군에서 약 60% 내외로 현저히 감소하는 것을 알 수 있었다. 본 실험 결과는 팽이버섯 추출물의 항암 활성을 확인 한 것으로 향후 이의 추출물을 이용한 세포신호전달과정 등에 관한 연구와 건강기능성식품의 소재로의 제품 개발에 유용하게 사용될 수 있음을 나타내는 주요한 자료로 판단된다.

블루베리 잎의 영양성분 분석 및 항산화 활성 (Nutritional Composition and in vitro Antioxidant Activities of Blueberry (Vaccinium ashei) Leaf)

  • 정희록;조유나;정지희;김현주;허호진
    • 한국식품저장유통학회지
    • /
    • 제19권4호
    • /
    • pp.604-610
    • /
    • 2012
  • 블루베리 잎의 무기질은 칼슘과 칼륨의 순으로 많았으며, 유리당은 glucose가 대부분을 차지하였다. 아미노산은 glutamic acid, aspartic acid, leucine의 순으로 많이 함유되어 있었으며, 지방산 함량은 eicosenoic acid, palmitic acid순으로 높았다. vitamin C와 total flavonoid 함량은 각각 $112.64{\pm}0.02$ mg%, $13.09{\pm}0.01$ mg%이었다. 블루베리 잎 80% 에탄올 추출물을 이용한 분획물의 total phenolics 함량은 에틸아세테이트 분획물이 50.51 mg of GAE/g로 제일 높았으며, 블루베리 잎 에틸아세테이트 분획물의 ABTS 라디칼 소거활성 및 FRAP assay는 농도 의존적으로 항산화 활성이 증가하는 경향을 보였다. 본 연구 결과를 종합해볼 때, 생리활성 소재로서의 phenolics를 함유한 블루베리 잎은 양질의 영양학적 성분 구성 및 in vitro 항산화 효과를 기초로 한 고부가가치 건강지향 식품 소재로서의 활용가치가 높다고 판단된다.

당귀 잎의 항산화 활성 (In vitro Antioxidant Activity of the Aqueous of Angelicae gigas Nakai Leaves)

  • 박성진;윤정한;김영언;윤원병;김종대
    • 한국식품저장유통학회지
    • /
    • 제18권6호
    • /
    • pp.817-823
    • /
    • 2011
  • Angelicae gigas Nakai has been used as a traditional medicine as well as an edible vegetable in South Korea. In this study, the total phenolic and flavonoid content and antioxidants of A. gigas Nakai leaves were examined in vitro via hydroxyl-radical-scavenging activity, reducing power activity, metal chelating assay, and DPPH-free -radical-scavenging assay. Among all the extracts from A. gigas Nakai leaves, the ethanol extract showed the strongest effects in all of the assays. The $EC_{50}$ values for the DPPH-radical-scavenging activities of ethanol, methanol, and water extracts were 31.47, 42.14, and $58.47{\mu}g/mL$, respectively. Among the extracts from A. gigas Kakai leaves, the ethanol extract had the highest levels of total phenolics ($7.84{\pm}1.46$ mg TAN/g) and total flavonoids ($4.23{\pm}0.03$ mg QE/g), which correlated strongly with the individual phenolic-compound (p-hydroxybenzoic acid, vanillin, and trans-ferulic acid) contents. The ethanol extract also showed stronger antioxidant activity than tocopherol in hydroxyl- radical-scavenging activity assay. These results indicate that the ethanol extract of A. gigas Kakai leaves possesses significant antioxidant properties, which suggests its great potential as a functional-food ingredient in the food industry.

Effect of Cutting Height on the Feed Value and Drying Rate of Rye (Secale cereale L.) Hay

  • Li, Yu Wei;Kim, Jong Geun
    • 한국초지조사료학회지
    • /
    • 제37권3호
    • /
    • pp.201-207
    • /
    • 2017
  • Hay-making is one of the most common forage preservation practices in livestock operations. The objective of hay-making is to minimize nutrient loss by shortening field drying time. Measuring the impacts of cutting height of forage crop is necessary to optimize hay production balancing yield and quality, in order to obtain substantial biomass increase through harvest of regrowth. This experiment was conducted to investigate the impact of cutting height of rye (Secale cereale L.) on drying rate and hay quality. Heading stage rye was harvested at 8cm or 15cm stubble heights. Hay was daily tedded at 09:00 and sampled at 09:00, 13:00 and 17:00 to determine moisture content (MC). After two month of preservation, CP (crude protein), ADF (acid detergent fiber), NDF (neutral detergent fiber), IVDMD (in vitro dry matter disappearance), TDN (total digestible nutrient), RFV (relative feed value), DM (dry matter) loss, visual scores and total fungi count were determined for estimation of hay quality. Cutting height at 15cm could enhance the drying rate and CP content (p<0.05), but also increases DM loss (p<0.05) compared to cutting at 8cm. Cutting heights did not affect ADF, NDF, IVDMD, TDN and RFV value (p>0.05). Visual scores of rye hays cutting at 8cm and 15cm, ranged from 83 to 85. Cutting at 8cm tended to maintain higher core bale temperature and fungal count than cutting at 15cm during preservation, but there was no significant difference.

Reproductive Biotechnologies for Improvement of Buffalo: The Current Status

  • Purohit, G.N.;Duggal, G.P.;Dadarwal, D.;Kumar, Dinesh;Yadav, R.C.;Vyas, S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제16권7호
    • /
    • pp.1071-1086
    • /
    • 2003
  • Reproductive biotechnologies continue to be developed for genetic improvement of both river and swamp buffalo. Although artificial insemination using frozen semen emerged some decades back, there are still considerable limitations. The major problem appears to be the lack of efficient methods for estrus detection and timely insemination. Controlled breeding experiments in the buffalo had been limited and similar to those applied in cattle. Studies on multiple ovulation and embryo transfer are essentially a replica of those in cattle, however with inherent problems such as lower number of primordial follicles on the buffalo ovary, poor fertility and seasonality of reproduction, lower population of antral follicles at all stages of the estrous cycle, poor endocrine status and a high incidence of deep atresia in ovarian follicles, the response in terms of transferable embryo recovery has remained low with 0.51 to 3.0 per donor and pregnancy rates between 15 to 30%. In vitro production of buffalo embryos is a valid alternative to recovery of embryos by superovulation. This aspect received considerable attention during the past decade, however the proportion of embryos that develops to the blastocyst stage is still around 25-30% and hence the in vitro culture procedures need substantial improvement. Embryo cryopreservation procedures for direct transfer post thaw need to be developed for bubaline embryos. Nuclear transfer and embryo cloning is a technique that has received attention in various species during recent years and can be of immense value in buffaloes as they have a low rate of embryo recoveries by both in vitro and in vivo procedures. Gender pre-selection, genome analysis, gene mapping and gene transfer are a few of the techniques that have been studied to a limited extent during recent years and are likely to be included in future studies on buffaloes. Very recently, reproductive biotechnologies have been applied to feral buffaloes as well, but the results obtained so far are modest. When fully exploited they can play an important role in the preservation of endangered species.