• 한국가축번식학회지
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• 제22권4호
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• pp.411-417
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• 1998

본 연구는 난관액과 oviductal conditioned medium 이 다정자 침입과 체외배발달에 미치는 효과를 규명하기 위하여 수행하였다. 배양액내 난관액과 oviductal conditioned medium 의 첨가는 다정자침입율과 난자내 침입한 평균정자수를 감소시켰다 (P＜0.05). 정자와 난관액 그리고 oviductal conditioned medium 과 l.5, 3, 4.5 시간 공배양후 첨체반응의 성적은 대조구에 비하여 증가하였다. 체외수정후 체외발달 배양액에 난관액이나 oviductal conditioned medium 을 첨가하여 192 시간동안 배양했을 때 상실배와 배반포배발달율이 난관액과 oviductal conditioned medium 첨가구에서 유의적으로 높았다 (P＜0.05). 이러한 연구결과는 난관액과 OVCM 등 난관 유래물질은 다정자침입율과 난자내 침입한 평균정자수를 감소시키며 체외발달율을 높인다고 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• 제9권6호
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• pp.315-322
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• 2005

In this study, the authors investigated whether death of vascular smooth muscle cell (VSMC) had a pathological pertinence. Conditioned media obtained from rat aorta smooth muscle cell (SMC) that were induced death by expressing FADD in the absence of tetracycline (FADD-SMC) triggered death of normal SMC. DNA fragmentation and caspase-3 activation were observed in dying SMC by conditioned media. FADD-SMC showed transcriptional activation of tumor necrosis factor $(TNF)-{\alpha}$. Conditioned medium contained $TNF-{\alpha}$, indicating secretion of the cytokine from dying FADD-SMC. It was investigated if secreted $TNF-{\alpha}$ was functional. Conditioned medium activated ERK and p38 MAPK pathways and induced MMP-9 expression, whereas depletion of the cytokine with its soluble receptor (sTNFR) remarkably inhibited induction of MMP-9 by conditioned medium. These findings suggest that $TNF-{\alpha}$ in conditioned medium seems to be active. Then, contribution of $TNF-{\alpha}$ on death-inducing activity of conditioned medium was examined. Depletion of $TNF-{\alpha}$ with soluble $TNF-{\alpha}$ receptor decreased the death activity of conditioned medium by 35%, suggesting that $TNF-{\alpha}$ play a partial role in the death activity. Boiling of medium almost completely abolished the death-inducing activity, suggesting that other heat labile death inducing proteins existed in conditioned medium. Taken together, these results indicate that SMC undergoing death could contribute to inflammation by expressing inflammatory cytokines and pathological complications by inducing death of neighboring cells.

• Journal of Veterinary Science
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• 제22권6호
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• pp.86.1-86.13
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• 2021

Background: Conditioned medium is the medium obtained from certain cultured cells and contained secretome from the cells. The secretome, which can be in the form of growth factors, cytokines, exosomes, or other proteins secreted by the cells, can induce the differentiation of cells that still have pluripotent or multipotent properties. Objectives: This study examined the effects of conditioned medium derived from E17 rat brain cells on cells with pluripotent properties. Methods: The conditioned medium used in this study originated from E17 rat brain cells. The CM was used to induce the differentiation of primary colonies of mice blastocysts. Primary colonies were stained with alkaline phosphatase to analyze the pluripotency. The morphological changes in the colonies were examined, and the colonies were stained with GFAP and Neu-N markers on days two and seven after adding the conditioned medium. Results: The conditioned medium could differentiate the primary colony, beginning with the formation of embryoid-body-like structure; round GFAP positive cells were identified. Finally, neuron-like cells testing positive for Neu-N were observed on the seventh day after adding the conditioned medium. Conclusions: Conditioned medium from different species, in this case, E17 rat brain cells, induced and promoted the differentiation of the primary colony from mice blastocysts into neuron-like cells. The addition of CM mediated neurite growth in the differentiation process.

• Journal of Life Science
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• 제12권1호
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• pp.22-26
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• 2002

CS21 lymphoma cells that preferentially metastasize to lymph nodes after s.c. inoculation into BALB/c mice were grown in vitro in the presence of CA12 stromal cells isolated from lymph nodes. To obtain fundamental data on identification and characterization of the soluble factor(s) produced by CA12 stromal cells, we investigated the biological profile of the conditioned medium produced by CAl2 stromal cells. CAl2 conditioned medium has no affinity with Con A. CAl2 conditioned medium is associated with the proliferation of splenic T- and thymic T-cells with-out adding mitogen, although the conditioned medium cannot induce the differentiation of thymocytes. Additionally, we showed that H-7, not HA-1004 inhibits CS21 cell proliferation. These results suggest that CAl2 conditioned medium has a specific soluble factor(s) produced by lymph node stromal cells.

• Journal of Microbiology and Biotechnology
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• 제8권1호
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• pp.74-80
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• 1998

CS-2l lymphoma cells that preferentially metastasize to lymph nodes after s.c. inoculation into BALB/c mice were grown in vitro in the presence of CA- 12 stromal cells isolated from lymph nodes. In order to obtain fundamental data on the identification and characterization of the soluble factors produced by CA-12 stromal cells, the conditioned medium of CA-12 stromal cells that inhibited apoptosis of CS-21 cells was examined. Various analytical treatments revealed that the soluble factors in CA-12 conditioned medium are very sensitive to heat treatment and trypsinization. Moreover CA-12 conditioned medium has an affinity with heparin but not with Con-A. In addition to these, the activity of CA-12 conditioned medium was blocked by H-7, a PKC inhibitor, but the conditioned medium could not induce the differentiation of thymocytes. We concluded that CA-12 conditioned medium contains stromal cell-derived apoptosis-inhibitory molecules that play an important role in proliferation of CS-2l cells by suppressing cell apoptosis.

• 한국동물학회지
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• 제27권3호
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• pp.151-164
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• 1984

근원세포의 융합에 관여하는 융합유도물질의 존재를 구명하기 위하여 계배의 근세포를 배양하면서, muscle-conditioned medium (MCM) 이 근원세포의 융합에 미치는 영향을 조사하고, 근원세포로부터 배양액 내로 방출되는 단백질을 분석하여 다음과 같은 결과를 얻었다. (1) MCM은 뚜렷한 융합촉진 효과를 나타냈으며, 이러한 효과는 첨가된 MCM의 농도가 층가함에 따라 증가하였다. (2) MCM의 융합촉진 효과는 주로 근원세포로부터 방출되는 융합유도물질에 의하여 일어나는 것으로 판단된다. (3) 배양근원세포로부터 분비되는 45,000달톤과 65,000달톤의 단백질이 융합 유도물질일 가능성이 높다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권5호
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• pp.350-354
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• 2000

The growth promotion of a Taxus brevifolia cell suspension culture was investigated using conditioning factors. The conditioning factors produced and secreted from cultured cells usually stimulate cell division and the production of secondary metabolites. Therefore, the effective incubation time for the optimal secretion of conditioning factors was firstly determined for the promotion of cell growth. Conditioned media obtained by cultivating for 2 and 5 days showed the promotion of initial cell growth during the early cell growth period. However, the positive effect of the conditioning factors on the initial cell growth did not continue because of the depletion of the medium nutrients. Accordingly, the addition of a carbon source to the conditioned medium prolonged the positive effect on the cell growth. The addition of sucrose to the conditioned medium resulted in the maximum cell density being reached 4 days earlier compared to the control group and an increased substrate yield.

• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.207-216
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• 1998

Certain types of somatic cells, as well as follicular cumulus cells associating with mammalian oocytes, are known to produce beneficial effects on in vitro fertilization and pre implantation development of mammalian eggs when they are present in oocyte culture medium. To investigate the nature of the effects of somatic cells, the resistance of mouse oocytes against chymotrypsin treatment was examined after culture within various cell conditioned media. When mouse oocytes matured for 17-18 hr in the presence of cumulus cells were treated with 1 % chymotrypsin, half of them remained still alive even after 240 min $(t_{50}>240.0)$. In contrast half of mouse oocytes cultured without cumulus cells underwent degeneration within 65.0 min $(t_{50}=65.0{\pm}13.2min)$ of the same treatment. To see if the effects were duc to the secretory products of cumulus cells, mouse cumulus cells were cultured for 20 hr in medium containing 0.4% BSA and the supernatant of culture medium (conditioned medium) was taken. After maturation in the cumulus cell conditioned medium, mouse oocytes exhibited $t_{50}=190.0{\pm}10.8$ min upon chymotrypsin treatment whereas half of oocytes cultured without conditioned medium degenerated within 25.5 min. Human granulosa cell conditioned medium gave similar effects such that oocytes matured in conditioned medium exhibited $t_{50}=183.3{\pm}19.1$ min while $t_50$ of control group oocytes was $60.0{\pm}6.8$ min, Oocytes matured in vero cell conditioned medium exhibited $t_{50}=196.7{\pm}8.8$ min. On the other hand, amniotic cell conditioned medium resulted in the chymotrypsin resistance of $t_{50}=80.0{\pm}8.4$ min which was not statistically different from the control value of $t_{50}=48.0{\pm}13.2$ min. Based upon these results, it is suggested that certain somatic cell types including cumulus cells might change the biochemical properties of mouse oocyte membrane during meiotic maturation as revealed by the enhanced resistance against chymotrypsin treatment. Such effects of somatic cells appear to be mediated via the secretory products rather than direct communication between somatic cells and oocytes.

• Journal of Plant Biology
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• 제28권2호
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• pp.141-148
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• 1985

당근(Daucus carota L.) 세포 배양시 무성배 발생에 미치는 세포의 크기, 세포의 밀도, conditioned medium및 배지의 pH효과를 구명하기 위하여 실험한 결과는 다음과 같다. 세포배양시 구상형 배의 발생은 배양 4일째 최대에 달했으며 그 이후부터는 심장형 및 어뢰형 배로 발달되었다. 세포의 밀도가 높을수록 배 발생수가 증가하였으며 MS배지보다는 conditioned medium에서 양호하였고 배의 발달도 촉진되었다. 세포의 크기가 작을수록 생체중 및 배의 형성수가 증가하였으며 9o$\mu\textrm{m}$이상의 세포에서는 현저히 억제되었다. 배지의 pH가 4.0 혹은 7.0일때는 세포의 건물중이 현저히 감소하였고 배의 발생도 억제되었다. 그러나 pH 6에서 건물중이 최대에 달하였고 배의 발생도 양호하였다.

• 한국가축번식학회지
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• 제17권1호
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• pp.69-74
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• 1993

This study was investigated to examine the effect of conditioned medium from bovine oviductal cell(BOEC) in the co-culture system with BOEC on in vitro development of in vitro produced bovine embryos. Oocyte-cumulus complexes were cultured for 24 hrs in TCM-199 supplemented with 10% fetal calf serum, 1$\mu\textrm{g}$/ml FSH and 21U hCG, 1$\mu\textrm{g}$/ml oestradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. In vitro fertilization was performed with epididymal sperm and heparin (10$\mu\textrm{g}$/ml, 15min.) or caffeine(2.5mM)-treated spermatozoa. Oocytes were incubated with 1$\times$106 spermatozoa/ml for 18 hrs and then cultured in various culture system for 7 days. The development rates of 16-cell or blastocyst stages were recorded on 4, 7 days, respectively, after incubating. The proportions ofembryonic development into molulae and blastocysts were higher in cumulus cell co-culture(23.4%) and BOEC co-culture(34.3%) than in M199-FCS(6.1%). Similarily, the development rates into molulae and blastocysts were significantly higher in BOEC-conditioned medium than those in M199-FCS. Therefore, it is suggested that BOEC co-culture and BOEC conditioned medium increase significantly the development of in vitro produced bovine embryos in in vitro system.