• Clinical and Experimental Reproductive Medicine
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• 제25권1호
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• pp.65-70
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• 1998

This study was conducted to investigate the effects of Tissue Culture Medium 199 (TCM) and Dulecco's Modified Eagle Medium (DMEM) on the blastulation and grade of human oocytes on Vero cells in vitro. A cohort of 79 and 93 oocytes in metaphase II stage were used in TCM 199 and DMEM respectively. No differences were found in the numer of oocytes showing two-pronuclei between TCM (82.3%) and DMEM (86.0%). The number of fertilized oocytes reaching the blastocyst was not significant in TCM (60.0%) and DMEM (63.1%). A total of 89 blastocysts were categorized into the four grades (BG1, BG2, BG3 and early) depending on their morphology. The number of embryos achieving the blastocyst grade 1 (BG1) was significantly higher (p<0.05) in DMEM (50.8%) than TCM (15.0%). It is concluded that cultured oocytes in DMEM with glutamine on Vero cells should be significantly increased BG1.

• 한국발생생물학회지:발생과생식
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• 제12권1호
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• pp.51-56
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• 2008

제브라피쉬 배아 유래세포의 최적 성장조건을 확립하기 위해 3종류의 배지 DMEM, K-NAC, D-NAC 그룹에서의 세포 성장률을 비교하였다. 실험 결과, 접종밀도에 따른 성장률의 경우, DMEM에 비해 K-NAC 그룹에서 초기 접종 효율이 높게 나타났으며, 후기 성장률은 DMEM 그룹에서 높게 나타났다. K-NAC, DMEM 그룹 모두 FBS 농도에 의한 성장차는 보이지 않았으며, 0.1% embryo extract를 첨가한 배지에서 효과는 낮게 나타났으나 1% trout serum 첨가한 경우 매우 높은 성장률을 보였다(p<0.05). $2-3{\times}10^5$ 밀도로 접종한 그룹에서는 유의차가 없었으나, $4{\sim}5{\times}10^5$ 밀도에서는 DMEM 그룹이 K-NAC 그룹보다 다소 높은 성장률을 보였다(p<0.1). DMEM과 D-NAC 그룹에서의 FBS 농도에 따른 성장률을 비교한 결과, FBS 농도에 따른 성장차는 유의하지 않았으나(p<0.05), D-NAC의 모든 실험군이 DMEM 그룹에 비해 높은 성장률을 보였다(p<0.05).

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.1-7
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• 2000

Objective: Mammalian embryos undergo changes of energy environment for transfer from oviduct to uterus. Also, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration (oviduct - glucose: 0.5 mM, pyruvate: 0.32 mM, lactate: 10.5 mM; uterus - goucose: 3.15 mM, pyruvate: 0.1mM, lactate: 5.87 mM, respectively). This study was conducted to examine the effect of these energy sources added in DMEM with glutamine on the mouse embryo development. Methods: There was used ICR female mouse. Two cell embryos of mouse are collected by method of 'flushing'. Flushing fluid was used Ham's F-10 added to 20% FBS. The collected 2 cell embryos were cultured in media such as Control (only DMEM), group A and B (DMEM supplemented with 0.5 mM and 3.15 mM glucose), and group C and D (DMEM supplemented with 0.1 mM and 0.32 mM pyruvate), and group E and F (DMEM supplemented with 5.87 mM and 10.5 mM lactate). All experimental media supplemented with 20% hFF, respectively. Pattern of embryo development was observed to interval at 24hr during 96hr. Results : The media with glutamine added glucose (group A: 51.0%; group B: 48.4%) was significantly (p<0.05) higher than other experimental group in development into the morula stage after 24 hr in culture, but not significantly different compared with control and the rate of development into the blastocyst was significantly (p<0.05) low in the both of pyruvate (group C: 7.9% group D: 6.8%) and lactate (group E: 7.1%, group F: 7.1%) treatment group after 48 hr in culture. Development into the blastocyst and hatched balstocyst after 72 hr in culture revealed similarly in control (81.9%) and glucose treatment group (group A: 83.3%, group B: 82.8%). However, development into the hatched and attached blastocyst after 96hr in culture revealed significantly (p<0.05) development in the glucose treatment group (group A: 82.3%, group B: 78.5%) than control (63.2%), and its of pyruvate (group C: 34.1%, group D: 34.1%) and lactate (group E: 25.9%, group F: 33.3%) treatment group were significantly (p<0.05) lower than control similar to previous observations. Conclusion : The glucose added to the DMEM with only glutamine, as energy source, was highly to the rate of development compared with control, but the other energy sources were not, synthetically. Above refer to, the human reproductive organ (oviduct, uterus) contains energy sources of different concentration. Thus, further studies are will examine continuously to effects by interaction of different energy sources in the mouse embryo development, and these results will provide to foundation on the human embryo culture.

• 약학회지
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• 제33권3호
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• pp.161-166
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• 1989

Effects of dammarane glycosides of Panax ginseng on primary cultured chicken embryonic skeletal muscle cells were studied by microscopic observation and determination of the activity of acetylcholinesterase. Muscle cells were prepared from the breast of 12-day-old chicken embryo and cultured with either a medium consisted of 87.5% Dulbecco's Modified Eagle Medium (DMEM), 10% horse serum and 2.5% chicken embryonic extract or a medium consisted of 90% DMEM and 10% horse serum. It was observed that dammarane glycosides of Panax ginseng seemed to show the tendency to stimulate the growth and the differentiation of the muscle cells cultured with a medium consisted of 90% DMEM and 10% horse serum under microscopic observation. The activity of acetylcholinesterase in the muscle cells cultured with a medium consisted of 90% DMEM and 10% horse serum was increased by dammarane glycosides of Panax ginseng.

• 한국수정란이식학회지
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• 제22권3호
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• pp.179-184
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• 2007

체세포의 배양 방법은 체세포 핵이식에 의한 형질 전환 돼지 생산에 있어서 중요한 요인 중 하나이다. 본 연구에서는 돼지 태아 섬유 아세포의 효율적인 배양 방법을 수려하였다. 돼지 태아 섬유 아세포는 임신 33일째 태아로부터 제조하였으며, 돼지 태아 섬유아세포의 증식을 혈청과 배지 종류별로 분석하였다. 그 결과, 15% ES screened FBS가 포함된 DMEM 배지에서의 배양은 15% FBS보다 세포수의 증가가 훨씬 더 빠르게 나타났다. 또한, 태아 섬유아 세포는 DMEM/F-12와 다르게 ES midified DMEM과 DMEM 배지에서 $7{\sim}8$번째 계대까지 증식이 유지되었다. 이러한 배양 조건에서 PGK-neo 벡터(pKJ2)를 돼지 태아 섬유아 세포에 도입한 다음 12일간 $300\;{\mu}g/ml$ G418이 포함된 배지에서 선별하여 colony를 얻을 수 있었다. 이러한 결과는 본 연구에 이용된 배양 시스템이 transgenic vector를 도입시킨 돼지 체세포의 screening에 이용될 수 있음을 보여주고 있다.

• 생명과학회지
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• 제24권12호
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• pp.1301-1307
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• 2014

지방유래줄기세포(adipose-derived stem cell; ADSC)는 조직재생을 위한 탁월한 수단으로 인정되고 있으나 세포증식속도가 느려 ADSC의 배양용 배지에는 대게 fetal bovine serum (FBS)이 첨가된다. FBS는 세포에 다양한 영양분을 공급하지만 세포의 기능에 영향을 미칠 수 있는 미 동정 물질도 많이 함유하고 있다. FBS에 의한 예상밖의 영향과 동물유래물질의 오염을 방지하기 위해 ADSC의 무혈청 배양법에 관한 연구가 광범위하게 이루어지고 있다. 본 연구에서는 ADSC세포에 SV40의 T항원 유전자를 도입하여 증식속도를 향상시킨 ADSC-T세포주의 효율적인 무혈청 배양법을 확립하기 위해 ADSC-T의 세포증식에 미치는 아미노산복합체, 비타민 복합체 및 여러가지 영양분 혼합물(B27)의 영향을 검토하였다. 그 결과, ADSC-T세포를 DMEM/F12 무혈청 배지에 현탁하여 plate에 주입하였을 때는 증식하지 않았으며 아미노산, 비타민 및 B27 영양소복합체는 증식촉진효과를 나타내지 않았다. 그러나 ADSC-T세포를 유혈청 DMEM배지로 24시간 배양 후 DMEM/F12 무혈청 배지로 교체하여 배양했을 때는 세포가 증식하였으며 이때, 비타민 복합체와 B27 영양소복합체는 증식촉진효과를 나타내었다. 또한 Stem pro 배지를 이용하면 ADSC-T의 무혈청 부유배양이 가능한 것으로 나타났다. ADSC-T세포는 분자량 70 kDa 부근의 단백질을 다량으로 분비하였으며, 성장인자 중에서 insulin-like growth factor (IGF)와 fibroblast growth factor basic (FGF basic)는 유혈청 배양보다 무혈청 배양에서 더 많이 분비되었다.

• 한국수정란이식학회지
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• 제27권3호
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• pp.189-192
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• 2012

Although embryonic stem cells (ESCs) or ES-like cells are reported from many mammalian species other than the mouse, the culture system for murine ESCs may not be suitable to the other species. Previously many other research groups have modified either human or mouse ESC culture systems for bovine ESC culture. In this study, we compared three different culture mediums consisting of DMEM, ${\alpha}$-MEM or KnockOut$^{TM}$-DMEM (KO), which are modified from human or mouse ESC culture system, for the generation of bovine ESCs. In this study, some pre-requisite events which are important for establishment and long-term propagation of ESCs such as inner cell mass (ICM) attachment on feeder cells, primary colony formation and sustainability after passaging. Once the ICM clumps attached on feeder cells, this was designated as passage 0. In regards to the rate of ICM attachment, ${\alpha}$-MEM was superior to the other systems. For primary colony formation, there was no difference between DMEM and ${\alpha}$-MEM whereas KO showed lower formation rate than the other groups. For passaging, the colonies were split into 2~4 pieces and passed every 5~6 days. From passage 1 to passage 3, DMEM system seemed to be appropriate for maintaining putative bovine ESCs. On the other hand, ${\alpha}$-MEM tended to be more suitable after passage 6. Although ${\alpha}$-MEM support to maintain a ES-like cell progenies to passage 15, all three culture systems which are modified from human or mouse ESC culture media failed to retain the propagation and long-term culture of putative bovine ESCs. Our findings imply that more optimized alternative culture system is required for establishing bovine ESC lines.

• 한국가금학회지
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• 제23권1호
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• pp.9-17
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• 1996

이 연구의 목적은 주입구의 위치에 따른 병아리 배아의 생존율을 서로 비교하고 가장 적합한 주입구 위치를 찾기 위하여 실시되었다. 멸균처리된 핀셋을 사용하여 난각의 첨단부와 둔단부 그리고 옆부분에 주입구를 각각 만들었다. 연구 결과, 둔단부에 주입구 (BE1)를 만든 수정란의 발생율이 가장 높았으나 내부난각막이 불투명하여 혈관내 미세주입이 어렵다. 따라서 본 연구에서는 첨단부에 주입구를 만든 다음 이 주입구를 통하여 약 2 $\mu$L의 DMEM 용액을 2.5일령된 배자의 혈관에 주입하였고, 주입부위의 출혈을 막기 위해 DMEM 용액을 주입한 후 공기방울을 넣은 결과 생존율이 약 17.0% 이었다. 따라서 이러한 주입구에 의한 방법은 생식세포가 조작된 germline chimera 또는 형질전환닭을 생산하는데 매우 유용한 시스템으로 이용될 수 있을 것이다.

• Clinical and Experimental Vaccine Research
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• 제12권4호
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• pp.328-336
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• 2023

Purpose: Human peripheral blood mononuclear cell (PBMC)-based in vitro systems can be of great value in the development and assessment of vaccines but require the right medium for optimal performance of the different cell types present. Here, we compare three commonly used media for their capacity to support innate and adaptive immune responses evoked in PBMCs by Toll-like receptor (TLR) ligands and whole inactivated virus (WIV) influenza vaccine. Materials and Methods: Human PBMCs were cultured for different periods of time in Roswell Park Memorial Institute (RPMI), Dulbecco's minimal essential medium (DMEM), or Iscove's modified DMEM (IMDM) supplemented with 10% fetal calf serum. The viability of the cells was monitored and their responses to TLR ligands and WIV were assessed. Results: With increasing days of incubation, the viability of PBMCs cultured in RPMI or IMDM was slightly higher than that of cells cultured in DMEM. Upon exposure of the PBMCs to TLR ligands and WIV, RPMI was superior to the other two media in terms of supporting the expression of genes related to innate immunity, such as the TLR adaptor protein gene MyD88 (myeloid differentiation factor 88), the interferon (IFN)-stimulated genes MxA (myxovirus resistance protein 1) and ISG56 (interferon-stimulated gene 56), and the leukocyte recruitment chemokine gene MCP1 (monocyte chemoattractant protein-1). RPMI also performed best with regard to the activation of antigen-presenting cells. As for adaptive immunity, when stimulated with WIV, PBMCs cultured in RPMI or IMDM contained higher numbers of IFNγ-producing T cells and secreted more immunoglobulin G than PBMCs cultured in DMEM. Conclusion: Taken together, among the different media assessed, RPMI was identified as the optimal medium for a human PBMC-based in vitro vaccine evaluation system.

• 한국치위생학회지
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• 제12권4호
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• pp.733-739
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• 2012

Objectives : To Compare the degree of survival rate of gingival fibroblasts, which is concerned with teeth adherence based on the type of avulsed tooth's storage solution. Methods : Different media gingival fibroblasts were stored in Dulbecco's modified Eagle's medium(DMEM), Hank's balanced salt solution(HBSS), milk, saline, and green tea in for 1, 2, 3 hours. And, MTT assay was conducted to compare survival rate of human gingival fibroblasts. Results : 1. The survival rate of gingival fibroblasts in DMEM and HBSS was higher than thoes in other storage media( Milk> Saline> Green tea). 2. The survival rate of gingival fibroblasts in milk, saline and green tea decreased as time passed. 3. Because of low osmotic pressure, green tea showed decrease of survival rate of gingival fibroblasts. Conclusion : DMEM and HBSS were the most effective storage media for gingival fibroblast. Among milk, saline, green tea, milk is most effective storage media for keeping gingival fibroblasts. Milk is recommended for storage media of avulsed tooth for keeping viability of cells.