• Title/Summary/Keyword: fetal bovine serum

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The Growth of Human Osteoblasts in Culture Dishes Made with Poly-glycolic Acid Containing Fetal Bovine Serum (우태아 혈청이 포함된 Poly-glycolic Acid 배양판에서 인간 조골세포의 성장)

  • Choi, Jae Won;Kim, Yong Ha;Moon, Young Mi;Kim, Yoen Jung;Choi, Sik Young
    • Archives of Plastic Surgery
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    • v.33 no.5
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    • pp.612-615
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    • 2006
  • Purpose: An ideal bony construct can be divided into two broad categories: (1) the design and fabrication of biodegradable, biomimetic scaffolds that provide correct signals to induce osteogenesis: (2) the identification of an ideal source of osteoprogenitor cells to seed onto the scaffold. We selected poly-glycolic acid as a synthetic scaffold among various scaffolds because of these properties. Meanwhile, culture medium is supplemented with fetal bovine serum(FBS): such serum contains essential elements such as proteins, hormones, growth factors and trace minerals. The composition of FBS can be ideal for various cell growth in vitro. We supposed that we could enhance bone growth at a fractured site if FBS was mixed with synthetic scaffold-PGA. Methods: We cultured human osteoblasts in five different prepared culture dishes made with FBS and PGA mixture. The mixtures contained different ratio of FBS, that is, 0, 1.5, 3, 7, and 10%. We cultured human osteoblasts for seven days and examined the growth and attachment of the cells at the 1st, 3rd, 5th, 7th days, respectively. Results: In the mixture of 0% FBS and PGA, the growth of the cells lasted for one day. In 1.5 and 3% FBS and PGA, the growth of the cells was examined at the 3rd day, then minimally declined at the 5th and 7th days. In 7% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. In 10% FBS and PGA, the growth of the cells lasted for 5 days, then declined at the 7th day. Staining status of the osteoblasts with alkaline phosphatase showed pale pink color in 0% FBS and PGA groups, but bright pink color in 1.5, 3, 7, 10% FBS and PGA groups, especially in 3%, 7%. Conclusion: In consequence, the growth of human osteoblast was higher in the mixture of FBS and PGA groups than in pure PGA ones. It is assumed that the mixture of FBS and PGA affects the proliferation of human osteoblasts.

Effect of Bovine Follicular Fluid and Hormones on In Vitro Oocyte Fertilization and Development of Bovine Embryos (소의 난포액과 호르몬이 난포란의 체외수정 및 체외발달에 미치는 영향)

  • 최양석;송상현;최창용;하란조;강다원;최상용;윤창현;박충생
    • Journal of Embryo Transfer
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    • v.12 no.2
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    • pp.181-188
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    • 1997
  • This study was carried out to determine the effect of bovine follicular fluid(bFF), hormones, and fetal bovine serum(FBS) supplemented in the medium on the in vitro fertilization and development of bovine embryos. The ovaries were obtained from a local abattoir and placed in physiological saline kept at 30~32˚C and brought to the laboratory within 3~4 hours. The oocytes and follicular fluid were collected by aspiration from visible follicles, and the oocytes of grades I on the basis of the morphology of cumulus cells attached and the homogeneity of cytoplasmic granules were selected and used for maturation. The basal media used for oocyte maturation, fertilization and embryo development in vitro were Ham' F-10, TALP and TCM-199, respectively. The hormones supplemented in maturation medium were consisted of 35 pg /ml FSH, 10 pg /ml LH and 1 pg/mi estradiol-l7$\beta$. The bFF collected from 5~9 mm follicles was centrifuged, filtered and inactivated by heat-treatment at 56˚C for 30 min. FBS also was inactivated with the same method and kept at -20˚C until use. The embryos were co-cultured with the monolayer of bovine oviductal epithelial cells at 39˚C under 5% $CO_2$ in air for 9 days. The results obtained were summarized as follows: The fertilization rate of oocytes was found 87.4% from 10% FBS and hormones treatment for IVM, and 37.1% of these TVF embryos were developed to blastocyst stage in 10% FBS groups. Compared with this control system, the fertilization rate was decreased significantly(P<0.05) in the maturation without either FBS or hormones. These IVF embryos were developed to morula stage at the similar rate, but to blastocyst at significantly(P<0.05) lower rate in the embryo culture with or without FBS supplementation. The fertilization rate(82.9%) in hormones and 10% inactivated bFF was similar with 10% FBS and hormone groups(87.4%), but decreased significantly(P<0.05) in 20 or 30% bFF (61.0 or 66.0%), respectively. In vitro developmental competence to blastocyst stage in 10% FBS and 20% inactivated bFF(37.1% and 31.4%) was higher than in 10 or 30% inactivated bFF(20.0 or 19.2%) or 10, 20 and 30% fresh bFF(19.1, 21.0 and 17.5%) The results indicated that the in vitro fertillzation and development rate of the embryos should be improved in 10% FBS or 20% inactivated culture system and 20% inactivated bFF might be available economically for bovine oocyte maturation and embryo culture instead of fetal bovine serum.

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Co-culture of In Vitro Matured and Fertilized Bovine Oocytes with Oviductal Epithelium (체외성숙, 체외수정 우 난포란의 Co-culture에 관한 연구)

  • 고광두;양부근;김정익
    • Korean Journal of Animal Reproduction
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    • v.14 no.1
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    • pp.50-56
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    • 1990
  • Bovine oocytes obtained from follicles(2~5mm) of ovaries after slaughter were cultured in TCM 199 medium with 10~20% heat-inactivated estrus cow serum(ECS) for 25~27 hr, at 39$^{\circ}C$ under 5% CO2 in air. At the end of culture period, some oocytes were stained with 1% acetoorcein and examined for the evidence of oocyte maturation. The remainder were used to assess the potential of in vitro fertilization(IVF) with frozen-thawed spermatozoa and subsequent development in media with or without bovine oviduct epithelial cell (BOEC) co-culture. The results obtained were summarized as follows ; 1. The maturation rate of oocyte in vitro in TCM 199 medium with 15% ECS group(76.3) was superior to 10% ECS group(68.3%) and 20% ECS group(64.5%). 2. The IVF rates of oocytes matured in vitro, and formation rate of male and female pronuclei were 63.6%(77/121) and 93.5%(72/77), respectively. The incidence of polyspermy was very low(2.4%). 3. Of 73 oocytes fertilized in vitro and cultured in TCM 199 medium with 10% fetal calf serum for 7 days, 41(56.3%) were cleaved over 2-cell and only 1(2.4%) was developed beyond the 16-cell stage. 4. Of 76 oocytes co-cultured with BOEC, 58(76.3%) were cleavaged and 23(39.7%) were developed to morula and blastocyst stage. The results of this study indicate that co-culture with BOEC deserved a positive effect on the IVF oocyte development through the 16-cell block.

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The Effects of the Mixture of Fetal Bovine Serum and Poly-glycolic acid in Rabbit Calvarial Model (Poly-glycolic Acid(PGA)와 우태아 혈청(Fetal bovine serum, FBS)의 혼합물이 가토에서 골형성에 미치는 영향)

  • Sung, Yong Duck;Kim, Yong Ha;Moon, Young Mi;Kim, Kap Joong;Kim, Yeon Jung;Choi, Sik Young
    • Archives of Plastic Surgery
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    • v.34 no.3
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    • pp.298-304
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    • 2007
  • Purpose: This study was undertaken to investigate the osteogenic induction potential of PGA & FBS mixture on a calvarial defect in the rabbit. Methods: Twenty New zealand white rabbit, weighing from 3.5-4kg were allocated into each of the three groups. Four 8 mm sized bone defects were made on the parietal bone by drilling. In group I, the bony defects were implanted with $50{\mu}m$ thickness film containing mixture of PGA and FBS. In group II, with PGA only film, & in group III, the bony defects were left with no implants. Results were evaluated by using morphologic change, radiographic study, biochemical study and histologic examination at 1 week (group I n=7, group II n=7, group III n=14), 2 weeks (group I n=6, group II n=6, group III n=12) and 3 weeks (group I n=7, group II n=7, group III n=14) following implantation. Results: In the morphologic & radiographic study, the formation and corticalization of callus were observed earlier in group I than in groups II and III (p < 0.05). In histological examination, group I showed more abundant and faster new bone formation than in group II and III. In biochemical analysis, group I displayed more activity than in group II and III. Group I also showed more abundant osteopontin, osteocalcin than groups II and III. Conclusion: In conclusion, the results demonstrate that the mixture of PGA and FBS has an effect on osteoblastic formation in the rabbit model. It is considered that further evaluation of long term results on resorption, immunologic tissue reaction and response of applied mixture in the human model will be needed.

Determination of the Optimal Concentration of Fetal Bovine Serum for the Growth of Two Insect Cell and Viruses (두 가지 곤충 세포주에 대한 배양 및 바이러스 증식을 위한 최적 FBS 농도 결정)

  • Lee, Jae-Kyung;Koo, Hyun-Na;Woo, Soo-Dong
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.319-324
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    • 2007
  • To determine the optimal concentration of fetal bovine serum (FBS) on the growth of insect cells and the multiplicity of viruses, the growth of cells (Sf21 and Bm5) and viruses were examined on the various concentrations of FBS. In view of the viability, growth speed, proliferation of cells and the amount of FBS, the most proper concentration for the cell culture were 7% and 5% for Sf21 and Bm5, respectively. The multiplicity of viruses at the various concentrations of FBS was similar in both cell lines at 5 days post-infection (p.i.). However, it differed significantly at 2 and 3 days p.i. The proper concentration of FBS were 10% and 3% for Sf21 at 2 and 3 days p.i., respectively, and 5% for Bm5 at both 2 and 3 days p.i. These results suggested that the optimal concentration of FBS should be determined according to the used cell lines and viruses for their optimum production.

Composition of a Medium for Serum-free Culture of an Adipose-derived Stem Cell Line Established with a Simian Virus 40 T Antigen (Simian virus 40의 T항원 도입으로 수립한 지방유래줄기세포주의 효율적인 무혈청 배양법 및 무혈청 배지조성)

  • Kim, Gyu Bin;Joo, Woo Hong;Kim, Dong Wan
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1301-1307
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    • 2014
  • Adipose-derived stem cells (ADSCs) are considered promising tools for tissue regeneration. However, ADSCs have very poor proliferation capacity. Therefore, fetal bovine serum (FBS) is generally added to the culture media of ADSCs. As FBS contains many uncharacterized components that may affect cellular functions, methods for serum-free cultures of ADSCs have been widely investigated. In this study, to develop an efficient method for a serum-free culture of ADSC-T, we used an ADSC line established by introducing the simian virus 40 (SV40) T gene into primary ADSCs. We then investigated the effect of amino acids, vitamins, and other components on the growth of ADSC-T. When the ADSC-T cells were plated with DMEM/F12 serum-free medium, the cells did not proliferate, and the mixture of amino acids, vitamins, and B27 supplement did not increase the growth of the cells. However, when the ADSC-T cells were provided with serum-free DMEM/F12 after they had been cultured with serum-supplemented DMEM for 24 h, the cells proliferated, and the vitamins and B27 supplement increased the cell growth. Stem-Pro serum-free medium also appeared to be useful as a suspension culture for the ADSC-T cells. The ADSC-T cells secreted large amounts of proteins of around 70 kDa. Insulin-like growth factor (IGF) and fibroblast growth factor basic (FGF basic) were secreted by ADSC-T in larger amounts in the serum-free culture than in the serum-supplemented culture.

Effects of Glucose and Glutamine Concentrations on Hybridoma Growth ann Anti-HBsAg MAb Production in Enriched Medium (농축배지에서 Glucose와 Glutamine 농도가 하이브리도마 증식과 간염표면항원에 대한 단일클론항체 생산에 미치는 영향)

  • 전복환;조의철김동일
    • KSBB Journal
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    • v.5 no.4
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    • pp.365-371
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    • 1990
  • To improve the growth of mouse hybridoma 2c3.1 secreting anti-Hepatitis B surface antigen monoclonal antibody (anti-HBsAg MAb), we had constructed an enriched medium and observed the effects of fetal bovine serum and serum-free supplements including human serum albumin, 'insulin and transferrin', and monoethanolamine. For further enhancement of growth, the concentrations of two major energy sources, glucose and glutamine, were strengthened with various ratios in the enriched medium. Maximum cell growth and monoclonal antibody production obtained in various ratios of glucose/glutamine with an inoculation concentration of 2$\times$105 cells/ml were 0.73$\times$106-4.62$\times$106 cells/ml and 65.1-422.6 $\mu\textrm{g}$/ml, respectively. Glutamine was round to be a major energy source and a limiting nutrient in comparison to glucose for 2c3.1 cell cultivation in enriched media with low serum.

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소태아혈청과 Macromolecules 가 소 난자의 성숙과 핵 이식 후 배 발달에 미치는 영향

  • 임기순;양병철;이상기;성환후;장원경
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.33-33
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    • 2002
  • 소난자 체외성숙에 있어 소 태아혈청 (fetal bovine serum: FBS) 첨가시 높은 성숙율을 보이며 핵 이식 후 배 발달율에 있어서도 높다. 하지만 배양액 내에 포함된 소 태아혈청은 수정란 이식 후 태아의 발육 중 및 태어난 직후에 발생되는 송아지에서 나타나는 몇몇 비정상적인 현상들의 원인인 것으로 보고되고 있다 (Walker 등, 1998). 본 실험은 성숙배양시 소 태아혈청과 이의 대체물질로서 BSA 또는 PVA 와 같은 macromolecules의 첨가가 소 난자의 성숙과 핵 이식 후 배 발달에 미치는 효과를 알아보기 위하여 실시하였다. (중략)

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Effect of Bovine Oviductal Epithelial Cell(BOEC) Conditioned Medium on In Vitro Development of Bovine Embryos Fertilized In Vitro (난관상피세포 Conditioned Medium이 체외수정된 소 수정란의 체외 발달에 미치는 영향)

  • 오종훈;김동훈;정형민;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.69-74
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    • 1993
  • This study was investigated to examine the effect of conditioned medium from bovine oviductal cell(BOEC) in the co-culture system with BOEC on in vitro development of in vitro produced bovine embryos. Oocyte-cumulus complexes were cultured for 24 hrs in TCM-199 supplemented with 10% fetal calf serum, 1$\mu\textrm{g}$/ml FSH and 21U hCG, 1$\mu\textrm{g}$/ml oestradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. In vitro fertilization was performed with epididymal sperm and heparin (10$\mu\textrm{g}$/ml, 15min.) or caffeine(2.5mM)-treated spermatozoa. Oocytes were incubated with 1$\times$106 spermatozoa/ml for 18 hrs and then cultured in various culture system for 7 days. The development rates of 16-cell or blastocyst stages were recorded on 4, 7 days, respectively, after incubating. The proportions ofembryonic development into molulae and blastocysts were higher in cumulus cell co-culture(23.4%) and BOEC co-culture(34.3%) than in M199-FCS(6.1%). Similarily, the development rates into molulae and blastocysts were significantly higher in BOEC-conditioned medium than those in M199-FCS. Therefore, it is suggested that BOEC co-culture and BOEC conditioned medium increase significantly the development of in vitro produced bovine embryos in in vitro system.

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