• Title/Summary/Keyword: free-serum

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Development of Serum Free Medium and Optimization of Porcine Rotavirus Vaccine Production

  • Ko, Yun-Mi;Kim, Myoung-Hwa;Kim, Min-Young;Jeong, Yeon-Ho
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.207-209
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    • 2005
  • 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 vaccine product. For porcine Rotavirus vaccine production, it is necessary to develop serum free medium which do not cause those problems. A new serum free medium was developed for porcine Rotavirus vaccine based on DMEM, and the performance of developed serum free medium was evaluated in terms of Vero cell growth and Rotavirus vaccine production. The cell density, gown in serum free medium developed, was similar with that in serum supplemented medium. Also, it was higher than that in other commercially available serum free medium. The productivity of Rotavirus vaccine using serum free medium developed and optimum production strategies will be also discussed.

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Diagnostic Significance of the Serum Thyroid Hormone Indicies in Various Thyroid Diseases (각종 갑상선질환에서 혈청 갑상선홀몬치의 진단적 의의)

  • Han, Bong-Heon;Ko, Suk-Man;Yoon, Sang-Ryong;Ro, Heung-Kyu
    • The Korean Journal of Nuclear Medicine
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    • v.14 no.2
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    • pp.45-51
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    • 1980
  • In an attempt to evaluate the diagnostic significance of the serum thyroid hormones in various thyroid function states, the author measured serum $T_3$ uptake, serum $T_3$, serum $T_4$, serum free $T_4$ and free $T_3$ index in 27 cases of normal subjects, 11 cases of hypothyroidism, 152 cases of euthyroidism and 81 cases of hyperthyroidism by the radioimmunoassay method. The results were as follows: 1. The ranges of serum thyroid hormones in normal subjects were, serum $T_3$ uptake; $27.4{\sim}42.1%$, serum $T_3;\;93{\sim}245ng/dl$, serum $T_4;\;4.08{\sim}12.9ug/dl$ and serum free $T_4;\;0.57{\sim}1.53ng/dl(M{\pm}2S.D.)$. 2. Free $T_4$ index and serum $T_4$ show relatively high diagnostic value in euthyroidism group, and serum $T_3\;and\;T_4$ in hypothyroidism group, while serum $T_3$, free $T_4\;and\;T_4$ show relatively high diagnostic value in hyperthyroidism group. 3. There were significant correlation between free $T_4$ index and serum $T_4$(r=0.68), and between free $T_4$ index and serum free $T_4$(r=0.67) in hyperthyroidism group.

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Effect of Serum and Serum Free Media on the Developmental Competence of OPU Derived Bovine IVP Embryo

  • Choi, Byung-Hyun;Park, Bun-Young;Kong, Rami;Son, Mi-Ju;Park, Chan-Sang;Shin, Nyeon-Hak;Cheon, Hye-Young;Yang, Yeong-Rok;Lee, Jin-Woo;Jin, Jong-In;Kong, Il-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.4
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    • pp.305-310
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    • 2019
  • Embryos produced with serum show the alterations in their ultrastructure, impaired compaction, abnormal blastulation, aberrant mRNA expression profiles and large calf syndrome with greater incidences of stillbirths and deaths after birth. The aim of the present study was to describe in vitro embryo production by analyzing embryo production, fetal production and pregnancy rate in free-serum medium. The OPU-IVP data used in this study from 2016. Approximately, sixteen cows (Hanwoo), which belonged to the Institute of Gyeongsang National University, were used. Two experimental group is used in this study. Serum groups were conducted in March to July and free-serum group was conducted in September to December. The recovered cumulus-oocyte complexes were morphologically classified to four grades based on the compaction of cumulus cells layers and homogeneity of the cytoplasm. The number of oocyte was significantly greater in serum groups than that in free-serum groups (29.61 ± 0.63 vs. 15.6 ± 0.62; p < 0.05). Between serum and free-serum groups indicate that average of 1st and 2nd grade oocytes were no difference (2.38 ± 1.67 vs. 2.38 ± 1.48; p > 0.05), but number of 3rd and 4th grade oocytes were greater in serum groups than that in free-serum groups (7.31 ± 7.64 vs. 5.60 ± 6.29; p < 0.05). Embryo cleaved competence was higher in rate in free-serum groups than that in serum groups (62.1% vs. 58.3; p < 0.05). However, blastocyst developmental rate was no difference between serum and free-serum groups (33.1% vs. 43.5%; p < 0.05). 986 recipients were used for embryo transfer. Pregnancy rate was indicated that between serum and free-serum group was no difference (54.6% vs. 56.3%; p < 0.05). In conclusion, we developed the free-serum system for production of in vitro bovine embryos in order to meet the developmental and qualitative requirements for large scale commercial use.

Effects of serum on lymphokines producing capabilities of CD8+ T cells (Serum이 CD8+ T cell의 lymphokine 생산양상의 변화에 미치는 영향)

  • Ryu, Si-yun;Yoon, Won-kee;Cho, Sung-whan;Kim, Moo-kang;Kim, Tae-hwan
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.441-446
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    • 1994
  • The responsiveness of $CD8^+$ T-cell subpopulation according to serum-containing and serum-free conditions were investigated. Cells are freshly isolated from spleen of mature adult BALB/C mice between 13-20 weeks of age. $CD8^+$ T cells in serum-free conditions produce small amounts of IL-2, while significant amounts of IFNr following activation when compared the results of serum-containing conditions. These data indicate that serum-derived factors may play an important role in the alternations of $CD8^+$ T cell responsiveness.

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Functional characterization of primary culture cells grown in hormonally defined, serum-free medium and serum-supplemented medium (호르몬 한정배지를 이용한 세포 초대배양계의 확립)

  • Han, Ho-jae;Kang, Ju-won;Park, Kwon-moo;Lee, Jang-hern;Yang, Il-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.551-563
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    • 1996
  • This study investigated the properties of primary cultured proximal tubule cells in hormonally defined(insulin, transferrin, and hydrocortisone), serum-free medium or 10% serum-supplemented medium. The growth rate of the primary cultured proximal tubule cells was lower in the hormonally defined, serum-free medium than in the 10% serum- supplemented medium(p < 0.05), while the activities of brush border marker enzymes, alkaline phosphatase(AP), leucine aminopeptidase(LAP), and y-glutamyl transpeptidase(${\gamma}$-GTP) were increased(p < 0.05). The activities of these enzymes, however, decreased with the lapse of incubation time to 50-70% after 6 days culture compared to those of the freshly-prepared proximal tubules. The enzymatic activities of the primary cultured proximal tubul cells on 6, 9, 12, and 15 days of culture were significantly increased in the hormonally defined, serum-free medium compared to the 10% serum-supplemented medium(p < 0.05). The functional differentiation of the primary culture was examined by observing multicellular domes of the confluent monolayer, which is indicative of transepithelial solute transport. The dome formation by the proximal tubule cultures occurred at a higher frequency in the hormonally defined, serum-free medium than in the 10% serum-supplemented medium(p < 0.05). Upon electron microscopic examination, an increased density of the brush border was observed in the hormonally defined, serum-free medium compared to the cells grown in 10% serum-supplemented medium. The activities of $Na^+$glucose cotransporter($^{14}C$-a-MG uptake), $Na^+$phosphate cotransportere($^{32}P$ uptake) and $Na^+$ transporter($^{22}Na^+$ uptake) in the brush border membrane, and of $Na^+/K^+$-ATPase($^{86}Rb$ uptake) in the basolateral membrane were significantly stimulated in the hormonally defined, serum-free medium than in 10% serum-supplemented medium(p < 0.05). In conclusion, the primary cultured proximal tubule cells grown in the hormonally defined, serum-free medium demonstrated a slower growth rate, but the functions of cell were enhanced.

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Effect to Testosterone on the Growth of Primary Rabbit Proximal Tubule Cells in Serum-Free Medium (Testosterone이 토끼 근위 세뇨관 상피세포의 성장에 미치는 영향)

  • Chu Min-Ho;Park Seung-Joon;Chang Joo-Ho;Jung Jee-Chang
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.85-93
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    • 1995
  • In order to examine the effect of testosterone of the cell growth, using a primary rabbit kidney proximal tubule cell culture system, we observed the effect of 3 growth factors and testosterone supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of testosterone showed a potentiation of the effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (>10 nM) of testosterone indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, testosterone caused to potentiate the growth of the cell. In the presence of hydrocortisone, testosterone also potentiated the grwoth of the proximal tubule cells. According to the Northern analysis, testosterone increased significantly the level of ${\beta}-actin$ mRNA in proximal tubular cells of rabbit kidney. Consequently we may suggest that growth stimulatory effect of testosterone on the primary rabbit kidney proximal tubule cell in serum-free and hormonally defined media ascribed to increase the synthesis of ${\beta}-actin$, which is an important protein consisting of cellular microfilament.

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Transfer, Cryopreservation and Production of Bovine Embryos Cultured in Serum-Free System (Serum-Free Medium에서 배양한 한우 배의 내동성과 이식)

  • Im, Y.-J.;Kim, J.-H.;Song, H.-B.;Jung, Y.-G.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.133-145
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    • 2004
  • Serum-contain is commoly used for the production of in vitro-derived bovine embryos. However, were biological activity of serum varies from lot to lot, time consuming to choose better serum with good quality and risks of virus, bacteria and mycoplasma infection. This study established serum-free culture systems of in vitro embryo development to efficiently obtain a large number of blastocysts from ovaries of Hanwoo and oocytes maturation, cell number, tlerance of cryopreservation. Secondly, serum-contain medium is suspected of contributing to the large calf size, dystocia, cersarean sections, calf mortality and confirmed these blastocysts are high quality in terms of cyotolerance, high rates of pregancy and normal birth. For these reasons, Culture media (IVMD101 and IVD101) designed specifically for the preimplantation bovine embryo are rather simplistic, being based on salt solutions with additional energy substrates and growth factors. An improved serum-free medium (IVMD101) was developed for bovine oocytes maturation in vitro. Proportions of embryos developing to the blastocyst stage cultured in both IVD101(32.4%) and IVD101(34.5%) serum-free media were higher than in TCM199+10% FBS(12.4%) serumcontaining medium. Futhermore, the cell numbers per blastosyst obtained in the serum-free media were superior to those of blastocysts developed in serum-supplemented medium. Also, cell numbers of blastocysts obtained in the serum-free media were similar with blastocysts derived in vivo. Survival rate blastocysts after 24 hr incubation after thawing, the blastocysts cultured in both IVD101(94.5%) and IVD101(95.8%) serum-free media were higher than in TCM199+10% FBS (52.5%) serum-containing medium. After 72 hr incubation after thawing, hatching rates of blastocysts developed in IVD101(78.4%) and IVMD101(83.7%) were sighnificantly higher than that developed in the serum-supplemented medium(32.0%). The pregnancy rates almost not different between fresh blastocysts(38.2%) and frozen blastocysts(34.9%). The results suggested that the improved serum-free media(IVMD101 and IVD101) offer several advantages over culture in serum-cotaining medium, including increased rates of blastocyst formation and high cel numbers. Additionally, the survival and hatching rates of embryos product in serum-free media after post-thaw culture were superior to those of embryos produced in the serum-containg medium and useful for the production of high quality bovine embryos for cryo-preservation. These improved serum-free media are beneficial not only for the study of the mechanisms of early embryogenesis but also for mass production of good quality embryos for embryo transfer, cloning and transgenesis.

Development of Serum-Free Medium for Mouse-mouse Hybridoma Part II. Hybridoma Culture using Developed Serum-Free Media (Hybridoma배양을 위한 무혈청 배지의 개발 제2부 : 무혈청 배지를 사용한 Hybridoma배양)

  • 제훈성;최차용
    • KSBB Journal
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    • v.8 no.1
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    • pp.28-35
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    • 1993
  • The serum free medium was developed and used for the suspension culture of mammalian cells. Although there were the problems of the longer lag time and the smaller maximum cell concentration achievable, the higher specific productivity as well as other advantages of the serum free medium can make it a more realistic alternative. The existence of a staggering period in glucose concentration vs. time profile in the batch culture can be a practical indicating signal for performing fed batch culture. The concentration dependence of the effects of the additives in the serum free medium as well as its economic feasibility was also tested.

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Production, Cryopreservation and Transfer of Bovine Embryos Cultured in Serum-Free Medium

  • Hoshi, Hiroyoshi
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.3-7
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    • 2002
  • In vitro embryo culture techniques provide significant contributions not only for a basic research of fertilization and early embryogenesis, but also for a low cost mass production of bovine embryos for transfer, embryo diagnosis, nuclear cloning and the production of transgenic cows. This presentation introduces newly developed serum-free media (IVD101 and IVMD101) that are effective far high yields of transferable embryos of excellent quality from in vitro-matured and fertilized oocytes. Both serum-free media are superior to a conventional serum-containing medium on the increased rates of blastocyst formation, post-thaw embryo viability, and pregnancy after transfer. Furthermore, reduced risks of calf mortality and large calf syndrome are also observed for the serum-free-derived embryos. Serum-derived embryos contain a large number of lipid droplets and immature mitochondria in their cytoplasm that may account for the lower production of transferable embryos and poor embryo quality.

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Effect of ${\beta}-Estradiol$ on the Growth of Primary Rabbit Proximal Tubule Cells in Serum-free Medium (${\beta}-Estradiol$이 토끼 근위 세뇨관 상피세포의 성장에 미치는 영향)

  • Park, Sang-Ho;Chung, Joo-Ho;Ko, Kye-Chang;Jung, Jee-Chang
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.73-83
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    • 1993
  • In order to examine the effect of ${\beta}-estradiol$ on the cell growth, using a primary rabbit kidney poximal tubule cell culture system. We investigated the effect of ${\beta}-estradiol$ on alpha 1 (IV) collagen and ${\beta}-actin$ mRNA levels from primary rabbit kidney cell cultures, and also the effects of 3 growth factors and ${\beta}-estradiol$ supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of ${\beta}-estradiol$ showed a sizable potentiation effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (> 10 nM) of estradiol indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, ${\beta}-estradiol$ caused to potentiate the growth of the cell. In the presence of hydrocortisone, ${\beta}-estradiol$ also potentiated the growth of the proximal tubule cells. According to the Northern analysis, ${\beta}-estradiol$ increased the level of ${\beta}-actin$ mRNA, although mRNA level of the alpha I(IV) collagen was not changed significantly.

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