• Title/Summary/Keyword: serum-free

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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.

Serum Thyroxine to Thyroxine-Binding Globulin Ratio in Pregnancy and Newborn (신부(娠婦)와 신생아(新生兒)에서 혈청(血淸)Thyroxine/Thyroxine-Binding Globulin 비(比))

  • Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.16 no.2
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    • pp.59-61
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    • 1982
  • To evaluate the diagnostic value of the ratio of serum tyroxine$(T_4)$/thyroxine-binding globulin (TBG) for the thyroid status in pregnancy and newborn serum thyroxine, TBG, triiodothyronine, and free thyroxine levels were radioimmunoassayed in normal pregnant women at each of the trimesters, and the calculated serum $T_4/TBG$ ratios were compared with other parameters such as $T_3/TBG$ ratio and free $T_4/TBG$ ratio and free $T_4/TBG$ ratio. Serum $T_4$ levels were elevated with the propotionate increase in TBG levels during pregnancy, leading to the nearly constant value of serum $T_4/TBG$ ratios as in normal non-pregnant controls. In contrast, serum $T_3/TBG$ and free $T_4/TBG$ ratios varied considerably during pregnancy. In newborn, $T_4$ levels were nearly not changed with compared non-pregnant control value and TBG levels were elevated. The results indicate that serum $T_4/TBG$ ratio is a better parameter than others in evaluating the thyroid status during pregnancy and but newborn is a no better.

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The Experimental Effects of Pomegranate on the Cerebral Blood Flow and Obesity in Rats (석류가 뇌혈류 및 비만에 미치는 실험적 효과)

  • Jeong, Hyun-Woo;Jeon, Byung-Gwan
    • The Korean Journal of Food And Nutrition
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    • v.20 no.2
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    • pp.143-149
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    • 2007
  • To examine the effects of pomegranate on cerebral blood flow and obesity, we observed regional cerebral blood flow in normal rats, as well as body weight, serum total cholesterol, serum HDL-cholesterol, serum LDL-cholesterol, serum triglycerides, serum total lipids, serum phospholipids and serum free fatty acid levels in rats fed a high fat diet. The results were as follows. Pomegranate seed extract significantly increased regional cerebral blood flow and the peel extract increased regional the cerebral blood flow in a dose dependent manner. In addition, the pomegranate extracts(seed, peel, juice) decreased body weight, serum total cholesterol, serum LDL-cholesterol, serum triglycerides, serum total lipids, serum phospholipids and serum free fatty acid levels, and increased serum HDL-cholesterol in the rats fed a high fat diet. More specially the seed extract significantly decreased serum total cholesterol, serum triglycerides and serum phospholipid levels.

Lipid Composition of total Serum Lipids obtained from Healthy Subjects and Patients with Hypercholesterolemia (정상인 및 고콜레스테롤혈증 환자의 혈청 지방질 조성)

  • Yoon, Tai-Heon;Lim, Kyung-Ja;Kim, Eul-Sang
    • Journal of Nutrition and Health
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    • v.14 no.3
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    • pp.117-123
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    • 1981
  • The lipid composition of total serum lipids were investigated by latroscan TH-10 TLC analyzer in 69 healthy subjects and 62 patients with hypercholesterolemia. Total serum lipids level in healthy subjects was higher in females than males. There was a increase in total serum lipids level with increasing total serum cholesterol level in patients with hypercholesterolemia, regardless sex. Total serum lipids obtained from healthy subjects and patients with hypercholesterolemia were separated into phospholipids, free cholesterol, free fatty acids, triglycerides and cholesterol esters. Increasing the level of total serum cholesterol in the patients resulted in a decrease in the level of phospholipids, which was a major lipid fraction, indicating that HDL level in the patients was also decreased. However, cholesterol esters, triglycerides and free cholesterol levels in the patients were higher than those found in healthy subjects. The FC/CE ratio for male patients was lower than for healthy males, while in female patients, this ratio was similar to that observed in healthy females.

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Strategies for Transmissible Gastroenteritis Virus(TGEV) Vaccine Production by Swine testicle cells

  • Kim, Min-Young;Jeong, Yeon-Ho;Kim, Myoung-Hwa;Ko, Yun-Mi
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.203-206
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    • 2005
  • TGE (Transmissible Gastroenteritis) caused by a virus belonging to family coronavirus, results in an acute infection of the small intestine of the pig. The optimum operation variables such as multiplicity of infection (MOI), infection time and harvest time were investigated for TGE vaccine production by immobilized ST(swine testicle) cells. In the culture supplemented with 5% serum, maximum virus titer of $1.2{\times}10^6pfu/ml$ was obtained at the conditions of 0.01 MOI, 2day infection time, and 1 day harvest time. Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final product. For these reasons, much attention has been focused on the development of serum-free media. A new serum-free media (SFM) has been developed in order to produce TGE vaccine of high quality with low cost. The performance of SFM developed was compared with other commercially available serum-free media and serum supplemented media in terms of virus productivity. The cultures with serum-free media showed higher titer than that with serum supplemented media. Among various serum-free media tested, CHO-S-SFMII showed highest virus titer.

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Chicken Insulin-Like Growth Factor-I Stimulates Protein Synthesis of Chicken Embryo Myoblasts Cultured in Serum-Free Medium

  • Kita, K.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.1
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    • pp.17-20
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    • 2001
  • The effect of chicken IGF-I on protein synthesis of chicken embryo myoblasts cultured in serum-free medium was examined. When myoblasts were expanded to approximate 20-30% of well, the medium was changed to the serum-free medium including 0, 2, 20, 200 or 2000 ng/ml of recombinant chicken IGF-I. The culture medium including 10% fetal calf serum (FCS) was used as positive control. After 1 day of incubation, protein synthesis was measured by the incorporation of [$^3H$]-L-leucine. Thereafter cells were continued to incubate for further 18 hours, and the radioactivity in the protein was measured as an index of protein synthesis. The values for protein synthesis cultured in the serum-free medium without chicken IGF-I or with 2000 ng/ml of chicken IGF-I were the lowest. Protein synthesis was elevated with increasing chicken IGF-I concentration from 0 to 20 ng/ml. The values for protein synthesis in the 20 ng/ml and 200 ng/ml IGF-I groups were about half of that of the FCS group. The present study revealed that the potency of chicken IGF-I at the levels of 20 to 200 ng/ml to stimulate myoblast protein synthesis was about half of that of 10% FCS.

Comparison of Two Different Serum-free Media for In Vitro Culture of Bovine Embryos

  • Kim, Se-Woong;Jung, Yeon-Gil;Park, Jong-Im;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.229-234
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    • 2014
  • The aim of the present study was to compare two different serum-free media, modified synthetic oviduct fluid (mSOF) and modified potassium simplex optimization medium (mKSOM) containing 20% RD (RPMI1640 + DMEM, 1:1 v/v) (RD-mKSOM), for in vitro culture (IVC) of bovine embryos. After in vitro maturation and fertilization, the presumptive zygotes were cultured in two different serum-free conditions for 7 days and 9 days to evaluate blastocyst formation and hatching, respectively. Serum supplemented conventional CR2 medium was used as control. After 7 day of culture, there was no significant difference in cleavage and blastocyst formation rates among three groups (mSOF, 59.3 and 30.1%; RD-mKSOM, 65.0 and 41.5%; control, 51.6 and 38.0%, respectively). Hatching rate was significantly higher in control (69.0%) than other experimental groups (mSOF, 22.0%; RD-mKSOM, 39.5%) (P<0.0001 and P<0.001, respectively). Although both serum-free conditions showed lower hatching rates than serum-added control, in serum-free groups, RD-mKSOM showed significantly higher hatching rate than mSOF (P<0.001). In addition, one-step using RD-mKSOM may facilitate IVC procedure than two-step culture system. In conclusion, the results indicate that one-step RD-mKSOM is more suitable defined culture system for IVC of bovine embryos than two-step mSOF.

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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Serum Free Medium Development for Recombinant Erythropoietin Production using Novel Cell Line (QT35) (QT35 세포주에서 제조합 에리스로포이에틴 생산을 위한 무혈청 배지의 개발)

  • 주형민;김병기;김선영;김태한;김태용
    • KSBB Journal
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    • v.13 no.3
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    • pp.295-302
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    • 1998
  • Human Erythropoietin (EPO) gene is cloned in quail fibrosarcoma cell, QT35. Because molecular weight of EPO is similar to that of serum albumin, cell culture with serum containing medium makes purification of EPO very difficult. Using fractional factorial study, we have developed serum free medium for the recombinant QT35 cell lines, QT N4D4 and QT SY-IMP, which have cloned EPO with glutamine synthetase (GS) gene amplification system and with puromycin selective marker, respectively. Among the seven frequently used medium components, fibronectin, BSA, and EGF were the most important for EPO production. However, sufficient fibronectin supplement to the medium did not make any good attachment of QT35 to culture plate over 3 days. Therefore, to maximize EPO production, we attempted a medium-shift at confluence from serum containing medium to serum free medium(QT SFM6). Using the medium-shift protocol with QT SFM6, nearly the same productivity of EPO was achieved comparing with that without medium-shift. This result was true in both QT35 cell lines in three types of culture, i.e. T flask, microcarrier and roller bottle cultures.

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Changes in Cell Cycle-related Genes Expression of Chineses Hamster Ovary Cells cultured using Serum-free Media

  • Seo, Sung-Keum;Park, Hong-Woo;Choe, Tae-Boo
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.153-158
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    • 2003
  • The genome-wide program of gene expression during the cell division cycle response to serum free media in chinese hamster ovary(CHO) cells was characterized using cDNA microarrays. Many transcripts of genes showed variation during the cell cycle. Characterization is critical step toward understanding the basic cell cycle processes and serum-free media role in CHO cells

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