• 한국가축번식학회지
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• 제22권3호
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• pp.229-235
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• 1998

본 실험은 마우스 초기배의 체외배양에 있어서 배양액 첨가제로서 기초배양액에 RPMI 1640 아미노산, MEM 비타민 및 사람의 성숙 난포액(hFF)을 각각 5가지의 농도로 구분하여 첨가했을 경우, 각 첨가제가 배 발달에 미치는 영향과 각 첨가제의 적정 첨가농도를 검토하고자 수행하였다. 그 결과를 요약하면 다음과 같다. 1. 배양액에 RPMI 1640 아미노산을 무첨가, 0.25%, 0.5%, 1% 및 2%의 농도로 첨가했을 때 배반포 발생율은 각각 35.5%, 54.5%, 65.4%, 48.2% 및 57.4%로 나타났으며, 0.5% 첨가구에서 유의하게 높았다. 2. 배양액에 MEM 비타민을 무첨가, 0.25%, 0.5%, 1% 및 2%의 농도로 첨가했을 때 배반포 발생율은 각각 12.8%, 22.4%, 31.1%, 21.9% 및 19.0%로 나타났으며, 0.5% 첨가구에서 유의하게 높았다. 3. 배양액에 hFF를 무첨가 2.5%, 5%, 10% 및 20%의 농도로 첨가했을 때 배반포 발생율은 각각 20.6%, 20.9%, 21.2%, 18.9% 및 29.4%로 나타났으며, 20% 첨가구에서 가장 높았다.

• 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.

• Parasites, Hosts and Diseases
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• 제50권4호
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• pp.309-315
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• 2012

Clonorchis sinensis is a biological carcinogen inducing human cholangiocarcinoma, and clonorchiasis is one of the important endemic infectious diseases in East Asia. The present study investigated survival longevity of C. sinensis adult worms in various in vitro conditions to find the best way of keeping the worms longer. The worms were maintained in 0.85% NaCl, 1${\times}$PBS, 1${\times}$Locke's solution, RPMI-1640, DMEM, and IMDM media, and in 1${\times}$Locke's solution with different supplements. All of the worms died within 3 and 7 days in 0.85% NaCl and 1${\times}$PBS, respectively, but survived up to 57 days in 1${\times}$Locke's solution. The worms lived for 106 days in DMEM, and 114 days in both RPMI-1640 and IMDM media. The survival rate in RPMI-1640 medium was the highest (50%) compared to that in DMEM ($20{\pm}10%$) and in IMDM ($33.3{\pm}25.2%$) after 3 months. The 1${\times}$Locke's solution with 0.005% bovine bile supplement showed increased duration of maximum survival from 42 days to 70 days. Higher concentration of bile supplements than 0.005% or addition of glucose were disadvantageous for the worm survival. The worms died rapidly in solutions containing L-aspartic acid, L-glutamic acid, and adenine compared to L-arginine, L-serine, and L-tryptophan. In conclusion, the 1${\times}$Locke's solution best supports the worms alive among inorganic solutions for 57 days, and the RPMI-1640 medium maintains living C. sinensis adults better and longer up to 114 days in vitro than other media.

• 대한수의학회지
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• 제38권2호
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• pp.359-365
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• 1998

This study was conducted to isolate the protozoan parasite Babesia gibsoni by intraerythrocytic culture method of micoraerophilous stationary phase(MASP) and evaluate the possibility of application for the detection of B gibsoni in canine babesiosis. Also, indirect fluorescent antibody test(IFAT) and thick blood smear(giemsa stain), direct light microscopy (DLM), as control diagnostic tests, were conducted to compare diagnostic effects between MASP, IFAT and DLM. The results obtained from this study were summarized as follows. The protozoan parasite B gibsoni multiplied in 24-well polystyrene plate containing 1.2ml of canine red blood cell suspension in RPMI 1640 medium(pH 7.0) which is contained 20~40% normal canine serum(NCS) under the MASP condition of 5% $CO_2$ and 95% air at $37^{\circ}C$ incubator. Under the above MASP culturing system the percentage of parasitized erythrocytes(PPE) after incubation for 9 days reached the peak. The levels of PPE in MASP culture were shown more higher by exchanging the medium at 24 hour intervals. The parasite were purely isolated from MASP culture of canine red blood cells collected from dogs(pit bullterrier) infected with B gibsoni naturally. Among the total of 83 heads of pit bullterrier blood samples the positive rate was 32 heads(38.5%) in DLM, 45 heads(54.2%) in IFAT and 42 heads(50.6) in MASP culture. In negative cases of IFAT and DLM the isolation rates of B gibsoni by MASP culture were 16 heads(42.1%) of 38 heads and 16 heads(28.6%)% of 56 heads, respectively. From this study it was suggested that MASP culture method by RPMI 1640 medium was a reliable and useful diagnostic test for the diagnosis of B gibsoni infections in canine babesiosis.

• 대한수의학회지
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• 제38권3호
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• pp.589-594
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• 1998

The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

• 대한수의학회지
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• 제38권1호
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• pp.107-117
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• 1998

The cultivation for development of Ascaris suum from the second-stage larvae($L_2$) embryonated egg and the third-stage of rat-derived larvae($L_3$) recovered from lung of rats were performed to use the screening test of anthelmintics in vitro. The preparations of larvae for cultivation were that the artificially-hatched $L_2$ incubated the embryonated eggs of Ascaris suum in 0.1% formalin solution at $25^{\circ}C$ for 28 days and the rat-derived larvae($L_3$) recovered from the lung of rat infected with the embryonated eggs of Ascaris suum on 7 days after infection(DAI). The cultivation for development of Ascaris suum from the embryonated eggs($L_2$) and the rat-derived larvae($L_3$) for 14 days in RPMI medium 1640(with 5% bovine calf serum) were as follows : 1. The sizes of the liberated larvae($L_2$) which were artificially hatched from embryonated eggs with glass beads(diameter 5mm) were $190{\sim}250{\mu}m$ on 1 days in culture(DIC). The second-stage larvae were molted into third-stage larvae(early $L_3$; $250{\sim}300{\mu}m$) and the features of these larvae were first observed such as cephalic cuticle, esophageal lumen and anus etc. on 5 DIC and the sizes of late third-stage larvae were $250{\sim}450{\mu}m$ on 10 DIC. The sizes of early fourth-stage larvae($L_4$) were $500{\sim}700{\mu}m$ and the features of these larvae were more pronounced in internal organs on 15 DIC. 2. The sizes of third-stage larvae($L_3$) recovered from the lung of rats were $1,340{\sim}1,370{\mu}m$ and the feartures of cephalic cuticle, esophageal lumen, intestine, rectum, anus were visualized by inverted microscope on 1 DIC. The fourth-stage larva($L_4$) completed by third ecdysis were recognizable and sizes of early fourth-stage larvae were developed as $1,400{\sim}2,200{\mu}m$ on 5 DIC. The sizes of middle fourth-stage of larva were $1,900{\sim}2,300{\mu}m$ and the thickened epithelial rectum was observed on 10 DIC. The rectum and anus of late fourth-stage larva($L_4$ $2,500{\sim}3,200{\mu}m$) had developed completely in RPMI medium 1640 on 15 DIC.

• 대한수의학회지
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• 제27권1호
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• pp.41-45
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• 1987

The present study was carried out to investigate the optimal conditions on blastogenesis of guinea pig lymphocytes. A microculture system in conjuction with a semiautomatic multiple sample harvester was used to study the in vitro optimal condition of guinea pig lymphocytes. Careful analysis of lymphocyte transformation to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogen determined optimal conditions as: (a) 10% fetal bovine serum in RPMI-1640 medium (b) 48-hour culture period.

• Journal of Microbiology and Biotechnology
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• 제2권2호
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• pp.141-151
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• 1992

Physiological changes of the murine hybridoma cell line $S3H5/\gamma{2bA2}$ during adaptation to RPMI 1640 medium with 1%(v/v) fetal bovine serum were characterized in terms of cell growth, antibody production, morphology, and metabolic quotients. Cells adapted to 1% serum medium in T-flasks became sensitive to shear induced by mechanical agitation and required at least 5% serum in the medium or spent medium for cell growth in spinner flasks, while cells adapted to 10% serum medium in T-flasks could grow in 1% serum medium in spinner flasks. Consequently, long-term adaptation to low serum media may not give the expected growth enhancement. After adaptation to 1% serum medium, changes in cell morphology were observed. The cells in 10% serum medium were uniform and circular, while cells in 1% medium were irregularly shaped. The DNA contents, which were measured by flow cytometry, were almost constant among the cells in the range of 1% to 10%. Further, no significant changes in energy metabolism and specific monoclonal antibody production rate were observed among these cells.

• 대한수의학회지
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• 제26권2호
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• pp.245-249
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• 1986

The present study has been carried out to investigate the optimal condition on the blastogenesis of guinea pig lymphocytes. A microculture system in conjunction with a semiautomatic multiple sample harvester(SAMSH) was used to study the in vito optimal condition of guinea pig lymphocytes. Data were presented to show many variables that are Involved in studying the responses of guinea pig lymphocyte in a microculture system to the stimulation of Concanavalin A(Con A) and lipopolysaccharide (LPS). Analysis indicated that the conditions for optimal Con A as measured by incorporation of $^3H$-TdR include : (1) use of RPMI-1640 as culture medium, (2) use of $6{\mu}g$ of Con A, per culture, (3) use of $1{\times}10^6$ cells per culture. Conditions for optimal stimulation with LPS mitogen were similar to those used for Con A.

• 대한수의학회지
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• 제31권1호
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• pp.49-53
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• 1991

The present study has been carried out to investigate the optimal condition on lymphocyte blastogenesis of rabbit lymphocytes, whole blood culture and microculture system in conjunction with a semiautomatic multiple sample harvester(SAMSH) was used to study the In vitro optimal condition of rabbit lymphocytes. Data were presented to show many variables that are involved in studying the phytohemagglutinin(PHA) and lipopolysaccharide(LPS) response of rabbit lymphocyte in a microculture system. Analysis indicated that the conditions for optimal PHA as measured by incorporation of $^3H$-TdR include: (1) use of RPMI-1640 as culture medium. (2) use of $6{\mu}g$ of PHA, per culture. (3) 48-hours culture period. Conditions for optimal stimulation with LPS mitogen were similar to those used for PHA.