• Title/Summary/Keyword: FBS1

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Effect of Mouse Leukemia Inhibitory Factor on the Development of In Vitro Produced Pig Embryos (돼지 체외수정란의 발달에 미치는 Mouse Leukemia Inhibitory Factor의 영향)

  • 엄상준;정형민;박진기;이장형;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.3
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    • pp.207-216
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    • 1994
  • 본 연구는 돼지 난포란으로부터 생산된 수정란의 체외발달에 미치는 아미노산, 우태아혈청 (FBS)과 Leukemia Inhibitry Factor의 영향을 조사하였다. 돼지난포란은 도살된 돼지의 난소로부터 회수하여 39$^{\circ}C$, 5% CO2 배양조건하에서 1$\mu\textrm{g}$/ml FSH-p, 1$\mu\textrm{g}$/ml Estradiol-17$\beta$와 10%FBS가 첨가된 TCM-199 배양액내에서 42시간동안 성숙시켰다. 사출된 정자의 수정능 획득은 45와 90% Percoll density gradient법을 통한 원심분리에 의해 얻었으며, 이들 수정능획득된 정자를 성숙된 난포란이 함유된 배양액에 3$\times$105/ml의 농도로 주입하여 10$\pm$1시간동안 배양함으로서 체외수정을 유도하였다. 수정된 난포란은 ; 1) 10% FBS가 함유된 TCM-199, DMEM, mKRB 또는 CR1aa 배양액, 2) 아미노산 또는 10% FBS가 첨가된 CR1 배양액, 3) STO 세포 또는 mLIF (1,000 unit/ml) 첨가된 CR1aa(10%FBS) 배양액, 4) mLIF (1,000 unit/ml)를 수정 직후 또는 8-세포기 이후에 첨가된 CR1aa(10%FBS)의 네가지 배양조건에서 각각 분리 배양하였다. 그결과 체외수정란의 배반포까지 발달율은 아미노산과 10%FBS가 포함된 CR1 배양액에서 다른 배양액에서보다 양호하였고, 특히 8-세포기 이후에 mouse LIF를 첨가한 CR1aa(10% FBS) 배양액에서는 다른 배양조건보다 현저히 높은 결과를 보였으며, 부화 배반포까지도 배발달을 유도할 수 있었다. 따라서 돼지수정란의 발달에 있어서 배양액내에 아미노산과 FBS 및 mouse LIF의 첨가는 효과가 있으며, 특히 8-세포기 이후에 있어서 mouse LIF의 첨가는 돼지의 수정란을 배반포 이후의 단계에까지 발달시킬 수 있었다.

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Developmental Potential of Bovine Nuclear Transfer Embryos Cultured in Serum-free Medium

  • Lee, S. K;Kim, D. H.;G. S. Im;B. C. Yang;Park, H. S.;W. K. Chang;Lee, H. T.
    • Korean Journal of Animal Reproduction
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    • v.27 no.4
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    • pp.339-347
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    • 2003
  • The purpose of this study was to investigate the development of bovine nuclear transfer (NT) embryos cultured in serum-free conditions. Bovine NT embryos cultured in various culture conditions were compared blastocyst development, total cell number and apoptosis using TUNEL assay. In experiment 1, blastocyst rates of NT embryos were significantly higher (P<0.01) in FBS (22.0%) and BSA (26.6%) groups than in PVA (6.3%) group. Total cell number was significantly higher in FBS (78.4$\pm$19.4) and BSA (90.9$\pm$29.1) groups than in PVA group (46.0$\pm$0.0). Apoptotic cell number was significantly fewer in FBS (3.1$\pm$1.4) and BSA (1.7$\pm$1.4) groups than in PVA group (7.0$\pm$20.0) However, all of results were not different between the FBS and BSA group. In experiment 2, blastocyst rates of NT embryos were significantly higher (P<0.05) in fatty acid free-BSA (FAF-BSA) group (26.8%) than in fraction V-BSA group (11.2%). Total cell number were somewhat higher in FAF-BSA group (89.8$\pm$30.7) than in fraction V-BSA group (88.1$\pm$19.3). Apoptotic cell number were somewhat fewer in FAF-BSA (1.7$\pm$1.5) group than in fraction V-BSA group (4.2$\pm$2.9). These findings suggest that serum free condition were effective for the in vitro development of bovine NT embryos. Therefore, we concluded that fatty acid free-BSA has beneficial effect in development bovine NT embryos and can be use as a serum substitute.

Silk Protein as a Fetal Bovine Serum Substitute for Animal Cell Culture

  • Jo, You-Young;Kweon, HaeYong;Ji, Sang Deok;Kim, Jong Gil;Kim, Kee Young
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.487-497
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    • 2019
  • Fetal Bovine Serum (FBS) is an essential substance added to animal cell culture medium. However, its composition is unclear causing problems such as development of an immune response when cultured cells are transplanted into the human body. In this study, silk sericin, silk fibroin, and hemolymph obtained from silkworms were added to the cell culture medium in order to determine if it can replace FBS. After establishment of the cell culture, cell proliferation and expression levels of cell growth-related genes were compared with those of control cells (cells cultured in the medium with 10% FBS). Results showed that the test group treated with silk fibroin extracted from a Korean silkworm variety, Kumokjam could replace 10% FBS. In addition, expression levels of cell growth related genes such as Fibronectin and TGF-β1 increased significantly in cells cultured using silk fibroin, depending on the concentration used in cell adhesion and cell proliferation [24]. To date, no studies have been conducted to find a replacement for FBS. Thus, this study was carried out to develop a substitute for FBS by using silkworm-derived alternatives such as silkworm hemolymph, silk sericin, and silk fibroin, which are cheap and have various physiological effects, cell promoting effects, and can be mass produced.

Evaluation of in vitro ruminal fermentation of ensiled fruit byproducts and their potential for feed use

  • Mousa, Shimaa A;Malik, Pradeep K.;Kolte, Atul P.;Bhatta, Raghavendra;Kasuga, Shigemitsu;Uyeno, Yutaka
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.103-109
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    • 2019
  • Objective: Ensiling of tannin-rich fruit byproducts (FB) involves quantitative and qualitative changes in the tannins, which would consequently change the rumen fermentation characteristics. This study aimed to evaluate whether ensiled FBs are effective in mitigating methane emission from ruminants by conducting in vitro assessments. Methods: Fruit byproducts (grape pomace, wild grape pomace, and persimmon skin) were collected and subjected to four-week ensiling by Lactobacillus buchneri inoculant. A defined feed component with or without FB samples (both fresh and ensiled material) were subjected to in vitro anaerobic culturing using rumen fluid sampled from beef cattle, and the fermentation parameters and microbial populations were monitored. Results: Reduced methane production and a proportional change in total volatile fatty acids (especially enhanced propionate proportion) was noted in bottles containing the FBs compared with that in the control (without FB). In addition, we found lower gene copy number of archaeal 16S rRNA and considerably higher levels of one of the major fibrolytic bacteria (Fibrobacter succinogenes) in the bottles containing FBs than in the control, particularly, when it was included in a forage-based feed. However, in the following cultivation experiment, we observed that FBs failed to exhibit a significant difference in methane production with or without polyethylene glycol, implying that tannins in the FBs may not be responsible for the mitigation of methane generation. Conclusion: The results of the in vitro cultivation experiments indicated that not only the composition but also ensiling of FBs affected rumen fermentation patterns and the degree of methane generation. This is primarily because of the compositional changes in the fibrous fraction during ensiling as well as the presence of readily fermented substrates, whereas tannins in these FBs seemed to have little effect on the ruminal fermentation kinetics.

Enhancement of Hybridoma Cell Growth and Anti-Hepatitis B Surface Antigen Monoclonal Antibody Production in Enriched Media with Low Serum (저혈청농축배지에서 세포성장 및 간염표면항원에 대한 단일클론항체 생산의 증가)

  • 전복환;조의철김동일백승복
    • KSBB Journal
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    • v.5 no.1
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    • pp.87-94
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    • 1990
  • Enhancement of hybridoma cell growth and monoclonal antibody(MAb) production by the addition of a small amount of serum into both serum-free medium and enriched medium was studied. The enriched medium was constructed by mixing a basal serum-free medium and a nutrient-fortified RPMI 1640 medium. It was supplemented with human serum albumin, insulin, transferrin, and monoethanolamine. It was found that addition of low concentration of serum with other serum-free supplements was favorable for growth of a mouse hybridoma 2c3.1 cells. The concentration of serum was determined to 0.5%. The maximum cell concentration obtained in this enriched medium supplemented with 0.5% fetal bovine serum (FBS) was $3.06{\times}10^6$ cells/ml and the concentration of secreted anti-Hepatitis surface antigen (antiHBsAg) MAb was $159.7{\mu\textrm{g}}\;/\;ml$ compared to $43{\mu\textrm{g}}\;/\;ml$ in RPMI 1640 medium with 10% FBS and $50{\mu\textrm{g}}\;/\;ml$ in previously-developed serum-free medium. The 2c3.1 cell growth and MAb production could be enhanced considerably by using the enriched medium supplemented with 0.5% FBS and serum-free supplements instead of RPMI 1640 medium or serum-free medium. The enhancement in MAb production in the enriched medium was more noticeable.

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Stimulation of Cell Growth by Erythropoietin in RAW264.7 Cells: Association with AP-1 Activation

  • Seong Seu-Run;Lee Jae-Woong;Lee Yong-Kyoung;Kim Tae-Il;Son Dong-Ju;Moon Dong-Cheol;Yun Young-Won;Yoon Do-Young;Hong Jin-Tae
    • Archives of Pharmacal Research
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    • v.29 no.3
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    • pp.218-223
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    • 2006
  • Erythropoietin (EPO), a hematopoietic factor, is required for normal erythrocyte developments, but it has been demonstrated to have many other functions, and its receptor is localized in other tissues. In the present study, we investigated whether EPO can promote other cell proliferation and possible molecular mechanisms. EPO restored the inhibition of the RAW264.7 and PC12 cell growth by fetal bovine serum (FBS) withdrawal in a dose dependent manner, but not that of other cell types tested. The restoring effect of EPO was completed when the RAW264.7 cells were cultured in the medium containing as low as 3% of FBS, and 10 U/mL EPO could replace FBS. The restoring effect of EPO in the RAW264.7 cells was associated with the increased of c-Fos and c-Jun expression as well as AP-1 activation. These data demonstrate that EPO can stimulate RAW264. 7 cell as well as PC12 cell growth even when the cells were cultured without FBS or in the presence of small amounts of FBS in the medium, and this stimulating effect is associated with the activation of AP-1 transcription factor.

Management of Gastrointestinal Foreign Bodies with Brief Review of the Guidelines

  • Kaan Demiroren
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.26 no.1
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    • pp.1-14
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    • 2023
  • Foreign body (FB) ingestion is a common health problem that affects children more than adults. According to gastroenterologists' guidelines, the management of FB ingestion differs slightly between adult and children. This review aimed to compile adult and children guidelines and establish an understandable association to reveal the requirements and timing of the endoscopic procedure, which is the most effective and least complicated technique for gastrointestinal FBs. Coins, pins, and chicken and fish bones have been the most commonly ingested FBs. However, with their increasing use in recent years, large batteries with lithiumion conversion, stronger magnets composed of rare earth metals, such as neodymium, and superabsorbent objects have become the most morbid and mortal, necessitating new management strategies. Although the approach to gastrointestinal FBs is controversial, with different treatment options available in different disciplines, many studies have demonstrated the efficacy and safety of endoscopic procedures. Many factors influence the timing of endoscopy, including the nature, size, and location of the ingested object and the patient's clinical condition.

Chromosomal Analysis of Hanwoo Embryos by In Vitro Culture Condition (한우 체외 수정란의 체외 배양 조건에 따른 염색체 분석)

  • Choi, S.H.;Cho, S.R.;Han, M.H.;Kim, H.J.;Choe, C.Y.;Son, D.S.;Chung, Y.G.;Kim, S.K.;Sohn, S.H.
    • Journal of Embryo Transfer
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    • v.22 no.2
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    • pp.137-141
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    • 2007
  • Antioxidants were well known to be essential supplements in the complex media and serve as a reservoir of oxygen. In this study, Hanwoo COCs (cumulus oocytes complexes) were matured and developed in L-cysteine-TCM199 and analyzed metaphase chromosome. Maturation rate of Hanwoo COCs were 73.4%, 94.6% in 0.1% PVA, 0.1 mM L-cysteine, respectively and showed significantly different between the treatments (p<0.05). Blastocyst formation were revealed 20.3%, 10.0% in 5% FBS+TCM199, 0.1 mM L-cysteine+1% BSA, respectively. There were no significant difference among treatment groups. Metaphase chromosome were showed 18.3%, 12.0% in 5% FBS-TCM199, 0.1 mM L-cysteine, respectively and analyzable chromosome were 6.1%, 4.0% and had no differences between the treated groups. In the case of in vitro developmental stages, metaphase chromosome were showed 18.3%, 12.0% in $4{\sim}16$ cells stage, 43.1%, 13.0% in morulae stage and 94.8%, 100.0% in blastocyst stage. These results suggested L-cysteine has beneficial role for in virto maturation and development in Hanwoo COCs.

Comparison of Dry Matter Intake, Digestibility, and Nitrogen Balance in Spotted Deer (Cervus nippon) fed Forest by-product Silage, Arrowroot (Pueraria thunvegiana) Silage and Oak Browse (Quercus aliena) Hay (꽃사슴에 있어서 육림부산물 사일리지, 칡 사일리지 및 갈잎건초 급여에 따른 건물채식량, 소화율 및 질소출납의 비교)

  • Jeon, B.T.;Kim, Y.H.;Lee, S.M.;Kim, K.H.;Moon, S.H.
    • Journal of Animal Science and Technology
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    • v.44 no.1
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    • pp.87-94
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    • 2002
  • A comparison of intake, digestibility and nitrogen balance in spotted deer(Cervus nippon) fed forest by-product silage(FBS), arrowroot silage(ARS) and oak leaf hay(OLH) was made to examine the feeding value of forest by-product silage. Dry matter digestibility of ARS was significantly higher(p$<$0.05) than that of OLH while crude protein digestibility was significantly higher(p$<$0.05) in ARS and FBS than in OLH. The digestibility of crude fiber was highest in FBS. Dry matter intake of OLH was significantly higher(p$<$0.05) than that of ARS, and there was no significant difference with FBS. Nitrogen intake was higher in OLH and ARS than in FBS, but there was no significant difference. Fecal nitrogen was highest(p$<$0.05) in OLH and urinary nitrogen was highest(p$<$0.05) in ARS. Retained nitrogen was highest in FBS, however, there was no significant difference among treatments. In Conclusion. FBS was estimated as a useful roughage source for deer, showing high digestibility, dry matter intake and nitrogen utilization.

Ex vivo Expansion of CD34+ Hematopoietic Cells from Cord Blood in Various Culture Environments (다양한 배양 조건에서 제대혈 유래 CD34+ 조혈 세포의 체외 증식)

  • Choi, Yong Woon;Oh, Duk Jae
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.73-80
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    • 2006
  • This study compared cell expansion and colony forming ability in human cord blood stem cells cultured ex vivo with two kinds of cytokine combinations, two kinds of media, presence or absence of fetal bovine serum (FBS) and two or three dimensional (2D or 3D) culture environments. Purified $CD34^+$ cells were cultured in the IMDM (Iscove's Modified Dulbecco's Medium) and SFM (Serum Free Medium) containing a cytokine cocktail-I (coc-I) (EPO, GMCSF, SCF, and IL-3) or a cytokine cocktail-II (coc-II) (TPO, G-CSF, SCF, IL-6, and Flt3/Flk-2 ligand) with or without FBS. Generally, higher cellular and clonogenic expansion were observed in the coc-I cytokine condition, compared to coc-II cytokine condition. 3D (Methocult) and 2D (IMDM + coc-I + FBS) conditions gave the greatest cell ($2,258{\pm}456$ fold) and CFU (BFU-E: $652{\pm}19$, CFU-GM: $520{\pm}58$, CFU-GEMM: $339{\pm}100$ fold) expansions, respectively. In aspect of medium, IMDM was better than SFM, except for coc-II condition without FBS. In conclusion, 'IMDM + coc-I + FBS' and 'IMDM + coc-I' were the best CFU expansions on the occasion of all culture conditions. FBS and 2D conditions had affirmative effect on CFU expansion, generally. These data might provide a variety of notions about ex vivo expansion of hematopoietic stem cells.