• Title/Summary/Keyword: 세포 수

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The Cultivation of Anchorage-Dependent Animal Cell, Vero-6, on Macroporous Collagen Microcarrier (다공성 콜라젠 미립담체를 이용한 부착성 동물세포 Vero-6의 배양)

  • 최연수;최태부박정극
    • KSBB Journal
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    • v.8 no.5
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    • pp.465-472
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    • 1993
  • The comparison of the capabilities of cell growth of four different kinds of commercially available microcarriers was carried out by culturing anchorage-dependent animal cells, Vero-6, in a spinner flask. Using 3 g/l of Cytodex 3, the maximum final cell density was about $1.4{\times}10^6$ cells/ml and increased up to $2.0{\times}10^6$ cells/ml by increasing microcarrier concentration up to 5 g/l. The macroporous collagen microcarriers, VX-100, informatrix, and Cultispher-G showed the final cell concentration of $4{\times}10^6$ cells/ml, $2.1{\times}10^6$ cells/ml, and $3.2{\times}10^6$ cells/ml, respectively at the microcarrier concentration of 5g/1. According to this result, VX-100 showed better cell growth than informatrix and cultispher-G and also showed about 2 fold increase in final cell density comparing to Cytodex 3 solid bead. When the intermittent bead-to-bead transfer technique was introduced in the culture using Cytodex 3 bead and cultispher-G, the result was very successful and the cells grew out very well. The recovered cells by dissolving collagen microcarrier using collagenase enzyme were mostly viable and grew out very well on the surface of the fresh microcarriers.

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Effects of Recombinant Human Epidermal Growth Factor on the Proliferationand Radiation Survival of Human Fibroblast Cell Lines in Vitro (재조합 표피성장인자가 방사선이 조사된 섬유아세포 증식에 미치는 영향)

  • Kim, Hyun-Sook;Kang, Ki-Mun;Lee, Sang-Wook;Na, Jae-Boem;Chai, Gyu-Young
    • Radiation Oncology Journal
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    • v.24 no.3
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    • pp.179-184
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    • 2006
  • [ $\underline{Purpose}$ ]: To explore the effect of recombinant human EGF on the proliferation and survival of human fibroblast cell lines following irradiation. $\underline{Materials\;and\;Methods}$: Fibroblast was originated human skin and primary cultured. The trypan blue stain assay and MTT assay were used to study the proliferative effects of EGF on human fibroblast cell lines in vitro. An incubation of fibroblasts with rhEGF for 24 hours immediately after irradiation was counted everyday. Cell cycle distributions were analyzed by FACS analysis. $\underline{Results}$: Number of fibroblast was significantly more increased rhEGF (1.0 nM, 10 nM, 100 nM, 1,000 nM) treated cell than control after 8 Gy irradiation. Most effective dose of rhEGF was at 160 nM. These survival differences were maintained at 1 week later. Proportion of S phase was significantly increased on rhEGF treated cells. $\underline{Conclusion}$: rhEGF cause increased fibroblast proliferation following irradiation. We expect that rhEGF was effective for radiation induced wound healing.

Response of Metastatic Cancer Cells to Thermal Changes in vitro (배양온도 변화에 대한 전이성 암세포의 반응)

  • Ahn, San-Gil;Kwon, Young-Ee;Choi, Ho-Soon;Kwon, Jung-Kyun;Yoo, Jin-Young;Kim, Jong-Ryong;Kim, Won-Kyu
    • Applied Microscopy
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    • v.37 no.4
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    • pp.239-248
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    • 2007
  • Alteration of temperature is one of cancer therapies. In general, severe hyperthermia(around $43^{\circ}C$) and hypothermia(around $18^{\circ}C$) trigger apoptosis through mitochondria, though the specific mechanism is still unknown. CC-t6 and GB-d1 cell lines, which were originally derived from human cholangiocarcinoma and gall bladder cancer, were established from a metastatic lymph node. To investigate the mechanism of metastatic cancer cell response to thermal stresses, hyperthermia($37^{\circ}C{\rightarrow}43^{\circ}C$) and hypothermia($37^{\circ}C{\rightarrow}17.4^{\circ}C$) were designed. Thermal stresses did not induce apoptosis but necrotic cell death. Any alterations of caspase-3, -9, cytochrome c, Bax, and Bcl-2 were not found in both hyperthermia and hypothermia exposed fells using western blot analysis. In the transmission electron microscopy, typical necrotic, but not apoptotic, changes were observed. These results suggest that temperature changes induce cell death through necrotic pathway in metastatic cancer in vitro, and it can be one of effective anticancer methods.

Effects of Transient Treatment with Rotenone, a Mitochondrial Inhibitor, on Mouse Subventricular Zone Neural Stem Cells (미토콘드리아 저해제인 rotenone의 일시적 처리가 쥐의 뇌실 하 영역 신경 줄기 세포에 미치는 영향)

  • Park, Ki-Youb;Kim, Man Su
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1329-1336
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    • 2019
  • Subventricular zone (SVZ) in the brain contains neural stem cells (NSCs) which self-renew and differentiate to neurons and glial cells during postnatal period and throughout adulthood. Since fate decision to either proliferation or differentiation has to respond to intracellular and extracellular conditions, many intrinsic and extrinsic factors are involved. Among them, mitochondria have been reported to participate in fate decision of NSCs. In our previous report, we showed that long-term treatment of a mitochondrial inhibitor rotenone greatly inhibited neurogenesis. In this study, we examined the effects of short-term treatment of rotenone on SVZ NSCs. We found that (1) even one-day treatment of rotenone significantly reduced neurogenesis and earlier time points seemed to be more sensitive to rotenone, (2) a number of Mash1+ transit amplifying cells was decreased by one-day treatment of rotenone, (3) short-term treatment of rotenone eliminated most of the differentiated Tuj1+ neurons and Olig2+ oligodendrocytes, while glial fibrillary acidic protein (GFAP)+ astrocytes were not affected, and (4) sulfiredoxin 1 (Srxn1) gene expression was increased after one-day treatment of rotenone, indicating activation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway. All these results confirm that functional mitochondria are necessary during differentiation to neurons or oligodendrocytes as well as maintenance of neurons after differentiation. Also, these data suggest that temporary exposure to mitochondrial inhibitor such as rotenone might have long-term effects on neurogenic potential of NSCs.

조력 T세포 인자에 의한 면역반응의 억제

  • 이종길
    • The Microorganisms and Industry
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    • v.16 no.3
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    • pp.20-25
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    • 1990
  • 면역계에는 조력T세포(helper T cell)와는 반대로 면역반응을 억제적으로 조절하는 것이 주 기능인 억제성 T세포(suppressor T cell)가 존배한다는 것이 아려져 있다. 이 억제성 T세포가 존재한다는 개념이 면역학및 임상의학에 준 영향은 매우 지대하여, 앨러지나 자가 면역질환 같은 질병은 억제성 T세포의 결핍으로, 그리고 면역 결핍증같은 질병은 T세포가 지나치게 활성화된 것으로 설명되기에 이르렀다. 한마디로 거의 모든 저하된 면역반응은 억제성 T세포와 연관하여 설명되었다. 그러나 억제성 T세포가 발견된지 근 20년이 되고 그에 관한 논문이 5000여편에 달하는 최근까지도 억제성 T세포 및 이들 세포가 만드는 억제성 T세포 인자에 대한 실체는 정확히 규명되지 못하고 있다. 오히려 억제성 T세포가 조력 T세포처럼 별개의 세포군으로 존재한다는 것을 부인하는 학자도 최근 나오게 되었다.(이 분야의 연구에서 나타난 문제점들이 정리되어 있음). 그러나 억제성 T세포의 존재를 부인하는 학자들도 T세포가 면역 반응의 억제에 관여한다는 사실, 적어도 그 현상 자체를 부인하지는 않았다. 다시말하면, 면역반응이 T세포에 의하여 억제되는 현상은 이미 수 천년의 논문을 통하여 입증되고 있다. 면역반응의 억제에 있어서 억제성 T세포및 억제성 T세포 인자의 역할은 앞으로의 연구를 통해 명확히 규명될 것이며, 정말 억제성 T세포가 있느냐 없느냐 하는 것은 이 총설의 주제가 아니다. 다만 여기서는 조력 T세포도 경우에 따라서는 면역반응의 억제에 관여한다는 최근의 연구결과를 간단히 소개하고자 한다.

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Characteristics of Recombinant CHO Cell Growth and Erythropoietin Production in Serum-Containing Media and Serum-Free Media (혈청배지와 무혈청배지에서의 재조합 CHO 세포 성장과 Erythropoietin 생산)

  • 변태호;전복환
    • KSBB Journal
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    • v.11 no.3
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    • pp.288-294
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    • 1996
  • We have investigated the characteristics of recombinant CHO cell growth and erythropoletin(EPO) production at different concentrations of serum and inoculation density. Cell growth and EPO production were increased with the increase of serum concentration and inoculation density. Enhancement of CHO cell growth and EPO production by medium exchange using serum-free medium at the growth phase of cells was studied. It was found that the exchange of culture medium with serum-free medium was favorable for growth of cells and production of EPO. The maximum number of cell and concentration of EPO obtained by exchanging culture medium were $6.2{\times}105cells/$\textrm{cm}^2$ and 7,470units/m1, respectively, compared to $2.1{\times}105cells/\textrm{cm}^2$ and 2,380units/m1 in serum-containing medium without medium exchange. It was observed that CHO cell growth was correlated with EPO production in serum-free media.

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Latent Infection and Reactivation of Human Cytomegalovirus from Human Monocyte THP-1 Cells (인체단핵세포주 THP-1세포에서 Human Cytomegalovirus의 잠복감염과 재활성화)

  • 윤상임;문명숙;이찬희
    • Korean Journal of Microbiology
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    • v.37 no.2
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    • pp.145-150
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    • 2001
  • Reactivation of human cytomegalovirus (HCMV) from latency is often fatal to immunocompromised individuals. To understand the effect of HCMV on human monocytes where HCMV establishes latency, two human monocyte cell lines at different stages in differentiation, THP-1 and HL-60 were infected with HCMV. While the viability and morphology of HL-60 cells were not significantly affected by HCMV, the viability of THP-1 cells was dramatically decreased by HCMV infection. THP-1 cells infected with HCMV became aggregated and adhered to the surface of culture dishes, probably due to the increased expression of adherence molecules CD11b on the infected THP-1 cells. THP-1 cells established a latent HCMV infection were induced to differentiate by treatment with TPA and hydrocortisone. Recovery of infectious HCMV from the culture supernatant of differentiated THP-1 cells was dependent on the time of induction of differentiation after HCMV infection. Thus, in vitro model of reactivation of HCMV from latently infected monocytes was established.

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Fine Needle Aspiration Cytology of Poorly Differentiated "Insular Carcinoma" of the Thyroid - A Case Report - (갑상선의 저분화 "도암종"의 세침흡인 세포검사 - 1례 보고 -)

  • Lee, Seung-Sook;Ha, Chang-Won;Cho, Kyung-Ja;Jang, Ja-June
    • The Korean Journal of Cytopathology
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    • v.5 no.1
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    • pp.35-40
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    • 1994
  • A case of poorly differentiated "insular" carcinoma of the thyroid is presented and discussed with references to the literature. Un fine needle aspiration cytology of our case, the aspirates were highly cellular with tumor cells appearing as dispersed isolated cells as well as in dense clusters and syncytial formations. Occasional microfollicles containing colloid were evident. Most of nuclei were fairly uniform with considerable variability and a few showed definite atypical features. Nuclear grooving was additional features. Necrotic debris was not seen. Our cytologic findings were correlated well with histology, confirming typical insular pattern of tumor cells with the presence of occasional pleomorphic cells and papillary carcinoma-like features. With much attention to cytological features of insular carcinoma, it would be possible to diagnose a preoperative suggestive diagnosis, even though not definitive.

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Effects of Acanthopanax senticosus herb-acupuncture on cell proliferation and nitric oxide synthase expression in dentate gyrus of alcohol-intoxicated rats (오가피(五加皮) 약침(藥針)이 알코올 중독(中毒) 흰쥐의 치상회(齒狀回)에서 신경세포생성(神經細胞生成) 및 NOS 발현(發顯)에 미치는 영향(影響))

  • Kim Jong-Chul;Lee Eun-Yong;Kim Ho-Hyun;Kim Ee-Hwa
    • Korean Journal of Acupuncture
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    • v.20 no.2
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    • pp.67-76
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    • 2003
  • 목적 : 오가피 약침이 알코올에 의해서 중독된 Sprague-Dawley(S-D)계 흰쥐의 해마 치상회에서 새로운 신경세포 생성 및 NOS발현에 미치는 영향을 조사하였다. 방법 : S-D계 흰쥐에 알코올을(2g/kg) 3일간 연속으로 투여한후, 5일간 인체의 중완혈에 상응하는 부위에 오가피 약침(30mg/kg) 치료를 시행하였다. 치료효과를 관찰하기 위해서 BrdU-면역조직화학 염색법 및 NADPH-d-조직화학염색법을 이용하였다. 결과 : 알코올 처치군에서는 BrdU-양성세포수 및 NADPH-양성세포수가 모두 정상군에 비해서 감소한 반면에 알코올 처치후 오가피 약침으로 치료한 군에서는 BrdU-양성세포수 및 NADPH-양성세포수 모두 알코올 처치군에 비해서 증가하였다. 결론 : 오가피 약침치료가 알코올에 의해서 중독된 S-D계 흰쥐 해마 치상회에서 새로운 신경세포의 생성을 증가시키는 것을 확인하였으며, 그 기전으로 산화질소가 관여할 것으로 사려된다.

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Comparison of Cellular Senescence Phenotype in Human Fibroblasts from New-born and Aged Donors. (신생아와 노인 유래 섬유아세포의 노화과정에서의 세포학적 성질의 비교)

  • Yi, Hye-Won;Hwang, Eun-Seong
    • Journal of Life Science
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    • v.18 no.3
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    • pp.344-349
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    • 2008
  • Normal somatic cells proliferate for a limited number of doublings in culture and then enter an irreversible growth-arrest state called replicative senescence. Replicative senescence has been believed a reason for the limited cellular turnover and deterioration of tissue function in aged animals. However, there is no experimental evidence supporting this assumption. Furthermore, cells from aged person have been poorly characterized with an exception of the cases of T cells. In this study, we examined cell biological changes occurring in replicative senescence of fibroblast strains originated from a new-born (NHF-NB) and a 87 year old man (NHF-87). NHF-87 (and the cells from a 75-year old) proliferated to smaller population doublings and with longer doubling times than NHF-NB did. At early passages, NHF-87 exhibited a low senescence-associated ${\beta}-Gal$ (SA ${\beta}-Gal$) activity and lipofuscin level, typical markers for cellular senescence. Furthermore, they maintained low levels of lysosome and reactive oxygen species (ROS). All of these levels increased dramatically in the late passage NHF-87 quite similarly as those in the late passaged NHF-NB did. These results indicate that most cells originated from the aged maintain a phenotype of the cells originated from new-born donors and undergo replicative senescence with the same kinetics as that of the cells from new-born. It is also indicated that not SA ${\beta}-gal$ activity but cell proliferation rate may be qualified as a biomarker for cells aged in vivo.