• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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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.

• 대한수의학회지
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• 제36권3호
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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.

• KSBB Journal
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• 제13권3호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제14권1호
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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.

• Journal of Animal Science and Technology
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• 제58권3호
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• pp.11.1-11.6
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• 2016

Background: This study was investigated the effects of culture media conditions on production of eggs fertilized in vitro of embryos from ovaries of high grade Korean native cow, Hanwoo. Methods: The IVMD 101 and IVF 100 were used for in vitro maturation of selected Hanwoo oocytes and In vitro embryo culture after in vitro fertilization, respectively. The IVMD 101 and IVD 101 were used for in vitro culture and completely free of serum. Results: The cleavage rates of 2-cell embryos in reference to Hanwoo oocytes were 86.7, 92.9, and 90.1 % in the control group, IVDM101 medium and IVD101 medium, respectively which indicates that the IVDM101 medium and IVD101 medium may result favorable outcomes. The in vitro development rates of blastocysts were 12.4, 38.4 and 32.4 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. For hatched blastocysts, it was 5.3, 33.9, and 28.6 % in the control group, serum free IVMD101 medium and IVD101 medium, respectively. Hence, more favorable results were expected for the hatched blastocysts in which the IVMD101 medium and IVD101 medium were used than the control group. Average cell numbers of blastocysts were 128.3, 165.7, and 163.6 in the groups of TCM-199 + 10 % FBS medium, IVMD 101 medium, and IVD 101 medium, respectively which clearly show that the IVMD 101 and IVD 101 medium consequence significantly higher cell numbers compared to the control group (i.e., TCM-199 +10 % FBS medium). Pregnancy rate after embryo transfer was 39.6 % when the serum free medium was used which is higher than that of the medium supplemented with serum (32.8 %). In addition, stillbirth rates were 4.9 % in the group of serum free medium whereas it was 13.6 % in the serum supplemented medium (13.6 %). Conclusions: Taken altogether, serum free media, the IVMD 101 and IVD 101 represented more favorable results in the embryo development rate of embryos, cell numbers of blastocyst, and pregnancy rate. Of note, the IVMD 101 medium showed better outcomes hence, it might be a better option for future applications for in vitro culture of bovine embryos.

• 대한약리학회지
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• 제31권1호
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• pp.85-93
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• 1995

Testosterone이 serum-free medium에서 배양한 토끼의 신장 근위세뇨관 상피세포의 세포성장과 기능에 미치는 영향을 관찰한 바 다음과 같은 결과를 얻었다. 1. 토끼의 신장 근위세뇨관 상피세포는 testosterone 1 nM의 농도에서 유의한 세포 성장 촉진 효과를 나타내었고, testosterone 10 nM이상의 농도에서는 세포성장이 억제되었다. 2. Testosterone은 serum-free medium에서 성장촉진인자의 하나인 hydrocortisone을 growth supplement로 넣어준 serum-free medium에서 토끼 신장의 근위세뇨관 상피세포의 성장을 촉진시키었다. 3. Testosterone은 hydrocortisone을 growth supplement로 넣어준 serum-free medium에서 토끼 신장의 근위세뇨관 상피세포의 성장을 촉진시키었다. 4. Testosterone은 Northern blot analysis에 의하여 확인한 토끼 신장의 근위 세뇨관 상피세포의 ${\beta}-actin$ mRNA level은 증가되었다. 이상의 결과로 미루어 보아, serum-free 그리고 hormonally defined media에서 testosterone이 토끼의 신장 근위세뇨관 상피세포의 성장 및 기능에 대하여 촉진적으로 작용하는 것은 cellular mecrofilament의 중요한 구성단백의 하나로 밝혀진 ${\beta}-actin$의 합성 증가에 기인하는 것으로 생각된다.

• KSBB Journal
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• 제5권1호
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• pp.87-94
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• 1990

본 논문에서는 10% fetal bovine serum(FBS)를 첨가한 RPMI 1640배지에 비하여 간염바이러스 표면항원에 대한 단일클론항체 생산에 있어서 효과적은 무혈청 배지에 낮은 온도의 혈청을 첨가하여 하이브리도마_2_c3.1 세포에서 그 효과를 조사하였다, 세포성장관 단일클론항체 생산을 증가시키기 위하여 기본 무혈청배지와 RPMI 1640 배지성분들의 농도를 균형있게 강화시킨 배지를 2:1(v / v)로 혼합하여 농축배지를 조성하고, 이 배지에 2mg / ml 인혈청 알부민 $5\;{\mu\textrm{g}}\;/\;ml$ insulin, $5\;{\mu\textrm{g}}\;/\;ml$ tran-sferrin, $10\;{\mu\textrm{g}}\;/\;ml$ monoethanolmine 등의 몇가지 무혈청 첨가물들을 첨가하였다. 이 농축 배지에 fetal bovine serum(FBS)과 supplemented bovine calf serum(sBCS)을 첨가하였을때의 세포성장과 단일클론항체 생산을 비교하여 FBS농도를 변화하여 세포성장과 단일클론항체생산의 증가를 시도하였다. 0.5%를 농축배지에 첨가함으로써 세포성장과 단일클론항체생산의 증가를 보았다. 최대 세포농도는 $3.06{\times}10^6$ cells / ml이었으며, 이때 생산된 단일클론항체는 10% FBS배지의 $43.0\;{\mu\textrm{g}}\;/\;ml$ 무혈청배지의 $50\;{\mu\textrm{g}}\;/\;ml$보다 3배이상 높은 $159.7\;{\mu\textrm{g}}\;/\;ml$이 생산되었다.

• KSBB Journal
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• 제8권1호
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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.

• 대한수의학회지
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• 제34권3호
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• pp.441-446
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• 1994

serum factor가 $CD8^+$ T cell의 lymphokine 생산양상에 미치는 영향을 알아보기 위하여 13-20주령의 BAL B/C 마우스로 부터 $CD8^+$ T cell를 분리한 후 serum-containing medium과 serum-free medium을 사용하여 배양하였다. serum-free medium에서 배양한 $CD8^+$ T cell이 분비하는 lymphokine의 양은 serum-containg medium에서의 결과와는 달리 IL-2의 생산양은 낮았으나 IFNr의 생산양은 상당히 높았다. 이와같은 결과로 미루어 serum-derived factor가 $CD8^+$ T cell의 lymphokine 생산에 영향을 미친다고 생각된다.

• 대한약리학회지
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• 제29권1호
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• pp.73-83
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• 1993

Steroid hormone의 하나인 ${\beta}-estradiol$이 serum-free medium에서 배양한 토끼의 신장 근위세뇨관 상피세포의 세포성장과 기능에 미치는 영향을 관찰한 바 다음과 같은 결과를 얻었다 1. Serum-free medium에서 토끼의 신장 근위세뇨관 상피세포는 ${\beta}-estradiol$ 1 nM의 농도에서 유의한 세포 성장 촉진 효과를 나타내었고, ${\beta}-estradiol$ 10 nM이상의 농도에서는 세포성장이 억제되었다. 2. ${\beta}-Estradiol$은 serum-free medium에서 성장촉진인자의 하나인 hydrocortisone을 뺀 조건하에서 세포 성장을 증가시키었다. 3. ${\beta}-Estradiol$은 hydrocortisone을 growth supplement로 넣어준 serum-free medium에서 토끼 신장의 근위세뇨관 상피세포의 성장을 촉진시키었다. 4. ${\beta}-Estradiol$은 Northern blot analysis에 의하여 확인한 alpha I (IV) collagen mRNA level에는 별다른 변화를 보이지 않으나, ${\beta}-actin$mRNA level은 증가되었다. 이상의 결과로 미루어 보아, serum-free 그리고 hormonally defined media에서 ${\beta}-estradiol이$ 토끼의 신장 근위세뇨관 상피세포의 성장 및 기능에 대하여 촉진적으로 작용하는 것은 cellular microfilament의 중요한 구성단백의 하나로 밝혀진 ${\beta}-actin$의 합성 증가에 기인하는 것으로 생각된다.