• 제목/요약/키워드: Colony forming efficiency assay

검색결과 11건 처리시간 0.018초

나노물질의 세포독성 평가법으로 Colony Forming Efficiency Assay에 대한 검증연구 (Pre-validation of Colony Forming Efficiency Assay for Assessing the Cytotoxicity of Nanomaterials)

  • 조은혜;이재우;박선영;김필제;최경희;엄익춘
    • 한국환경보건학회지
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    • 제41권1호
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    • pp.17-23
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    • 2015
  • Objectives: The cytotoxcities of Au, Ag, SWCNT, $SiO_2$, and ZnO nanomaterials were evaluated in order to assess their potential toxicological effects in in vitro cell models using colony forming efficiency (CFE) assay. Methods: The CFE assay of the test materials was carried out on Hep G2 cells. The size distribution of nanomaterials was studied by transmission electron microscopy (TEM). Changes in cell viability after treatment with a toxicant will result in a decreased number of colonies formed in comparison to solvent. Results: The TEM images show that all the particles except SWCNT and ZnO can be considered approximately spherical. The gold and $SiO_2$ nanoparticles show no response (no toxicity) in concentration response experiments. A statistically significant toxic effect was found in Hep G2 cells treated with Ag, SWCNT and ZnO nanomaterials. Conclusion: In this study, we considered CFE assay to be a promising test for screening studies for cytotoxicity with physicochemical analysis.

자기 활성 세포 분리법과 군체 분리법으로 분리된 건 줄기세포의 자가 재생 능력 및 분화능 효율 비교 (Comparison of Efficiency of Self-renewal and Differentiation Potential in Tendon-derived Mesenchymal Stem Cells Isolated by Magnetic-activated Cell Sorting Method or Colony Picking Method)

  • 이모세;최유림;윤동석;이진우;윤길성;최우진;한승환
    • 대한족부족관절학회지
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    • 제18권3호
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    • pp.100-107
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    • 2014
  • Purpose: The purpose of this study is to evaluate the efficacy of mesenchymal stem cell (MSC) isolation by the magnetic-activated cell sorting (MACS) method in tendon tissue-derived cells compared to the colony picking method for isolation of MSCs by picking colony-forming cells. Materials and Methods: Human tendon-derived cells were isolated by enzyme digestion using normal tendon tissues from three donors. We used the magnetic kit and well-known MSC markers (CD90 or CD105) to isolate MSCs in tendon-derived cells using MACS. Cloning cylinders were used to isolate colony-forming cells having MSC characteristics in tendon-derived cells. Colony-forming unit-fibroblast (CFU-F) assay was used to evaluate the self-renewal capacity of cells isolated using the colony picking method or MACS. For comparison of differentiation potentials into osteogenic or adipogenic lineage between two groups, alizarin red S and oil red O staining were performed at 14 days after induction of differentiation in vitro. Results: Flow cytometry results showed that early passage tendon-derived cells expressed CD44 in 99.13%, CD90 in 56.51%, and CD105 in 86.19%. In the CFU-F assay, CD90+ or CD105+ cells isolated with MACS showed larger colony formation in size than cells isolated using the colony picking method. We also observed that CD90+ or CD105+ cells were constantly differentiated into both osteogenic and adipogenic lineages in cells from all donors, whereas cells isolated using the colony picking method were heterogeneous in differentiation potentials to the osteogenic and adipogenic lineages. Conclusion: CD90+ or CD105+ cells isolated using MACS showed superior MSC characteristics in the self-renewal and multi-differentiation capacities compared with cells isolated using the colony picking method.

파두가대황(巴豆加大黃)이 항종양작용(抗腫瘍作用)과 자연살해세포(自然殺害細胞)의 활성(活性)에 미치는 영향에 대한 실험적 연구

  • 노훈정;전병훈;문구;문석재
    • 대한한방종양학회지
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    • 제2권1호
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    • pp.75-90
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    • 1996
  • This experiment was designed to study the antitumor effects and Activity of Natural Killer Cell of semen Tiglii plus Rhizoma Rhei. The cytotoxic and antitumor effects were evaluated on human cell lines(A549, Caki-1, LL2, Sarcoma 180, NIH/3T3) after exposure to prebrewed Semen Tiglii plus Rhizoma Rhei water extract 0.1, 0.2, 0.4, 0.8, 1.6mg/ml using in MTT assay, LDH, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. From the result of MTT assay, the cytotoxicity of ST(生巴豆霜), ST+RR(生巴豆霜加大黃) were concentration-dependently increased in both group of the ST and ST+RR, the cytotoxicity of ST+RR(生巴豆霜加大黃) was similar to that of ST(生巴豆霜). 2. From the result of LDH, the cytotoxicity of ST, ST +RR were concentrati -on-dependently increased in both group of the ST and ST+RR, the cytotoxicity of ST+RR was similar to that of ST. 3. The antitumor effect on A549 tumor cell from the result of colony forming efficiency showed the inhibitory effect on the growth in both group of the ST and ST+RR, the inhibitory effect on growth was low slightly in the ST+RR. 4. From the result of SRB assay, the antitumor effect on caki-1 tumor cell of ST, ST+RR showed the inhibitory effect on the growth in both group of the ST and ST+RR, the antitumor effect of ST+RR was similar to that of ST. 5. Median survival time and increased life span were increased slightly in both group of the ST and ST+RR. 6. The inhibitory effect on the growth of Sarcoma 180 and Lewis lung carcinoma tumor cell were increased slightly in both group of the ST and ST+RR. 7. The activity of NK cell was increased in the ST+RR.

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수치파두(修治巴豆) 및 파두가황연(巴豆加黃連)의 세포독성(細胞毒性)과 항종양(抗腫瘍) 효과(效果)에 관(關)한 실험적(實驗的) 연구(硏究) (Experimental Studies on the Change of Cytotoxic and Antitumor Effects according to the Prebrewed Method of Semen Tiglii and Rhizoma Coptidis)

  • 조성각;문구;문석재
    • 대한한방종양학회지
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    • 제1권1호
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    • pp.191-211
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    • 1995
  • This experiment was designed to study the change of cytotoxic and antitumor effects according to the prebrewed method of Semen Tiglii and Rhizoma Coptidis. The cytotoxic and antitumor effects were evaluated on human cell lines(A 549, Caki-1, LL2, Sarcoma 180, NIH/3T3) after exposure to prebrewed Semen Tiglii and Rhizoma Coptidis water extract 0.1, 0.2, 0.4, 0.8, 1.6 mg/ml using in MTT assay, LDH, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. The cytotoxicity from the result of MTT assay was low slightly in the ST II(炒巴豆霜), high in the ST III(醋炒巴豆). The cytotoxicity of ST I + RC(生巴豆霜加黃連) was similar to that of STI(生巴豆霜). 2. The cytotoxicity from the result of LDH was low slightly in the ST Ⅱ (炒巴豆霜), high in the ST III(醋炒巴豆). The cytotoxicity of ST I + RC(生巴豆霜加黃連) was similar to that of ST I(生巴豆霜). 3. The antitumor affect on A 549 tumor cell from the result of colony forming efficiency was low slightly in the ST II (炒巴豆霜) and ST I + RC(生巴豆霜加黃連). 4. The antitumor effect on Caki-1 tumor cell from the result of SRB assay was low slightly in the ST II (炒巴豆霜). 5. Median survival time and Increased life span increased slightly in the ST I RC(生巴豆霜加黃連) and ST II (炒巴豆霜). 6. The inhibitory effect on the growth of Sarcoma 180 and Lewis lung carcinoma tumor cell increased slightly in the ST I + RC(生巴豆霜加黃連) and ST II (炒巴豆霜).

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가미하고초산(加味夏枯草散)이 항암제(抗癌劑)의 항종양효과(抗腫瘍效果)와 종양세포(腫瘍細胞)에 미치는 영향(影響) (Influence of Gamihagochosan on The Antitumor effect of Anticancer Drug and The Proliferation of Tumor Cell Lines)

  • 김균택;전병훈
    • 대한한방내과학회지
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    • 제18권1호
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    • pp.175-190
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    • 1997
  • In order to investigate the effects of Gamihagochosan Extract(加味夏枯草散抽出液) on antitumor effects after human cell lines (A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurement of antitumor effect of Mitomycin C(MMC) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for the measurement of antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows : 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 Cells after exposure to the extract of Gamihagochosan extract depressed the growth of tumor cells by concentration of Garnihagochosan. 2. Antitumor activity of the ethanol extract from Gamihggochosan extract and MMC on ascites form of Ehrlich carcinoma in mice is slightly improved. Especially the mean of survival times in the group of 200mg/kg and MMC 0.1mg/kg is improved over 34.9%. 3. When Gamihggochosan extract and MMC are administered together, the weight of tumor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Gamihagochosan extract and MMC are more significantly improved than MMC alone. According to the above result, it could be suggested that Gamihagochosan extract has indirect antitumor effect by the increase of MMC uptake.

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죽엽석고탕가감방(竹葉石膏湯加減方)이 항암화학요법제(抗癌化學療法劑)의 세포독성(細胞毒性)과 종양세포(腫瘍細胞)의 성장억제(成長抑制)에 미치는 효과(效果)에 대한 연구 (Effects of Jukyeopseokgotanggagambang on Anti-tumor Chemotherapeutic Cytotoxicity and Lysosomal Enzymes of Tumor Cell)

  • 전승훈;문구;전병훈
    • 대한한방내과학회지
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    • 제18권1호
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    • pp.191-206
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    • 1997
  • In order to investingate the effects of Jukyeopseokgotanggagambang Extract on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI 1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. According to the change of colony-forming efficiency and SRB assay of Caki-1 cell, hep3B and A549 cells after exposure to the extract of Jukyeopseokgotanggagambang extract, that extract depressed the growth of tumor cells depending on its concentration. 2. Antitumor activities of the ethanol extract from Jukyeopseokgotanggagambang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of Jukyeopseokgotanggagambang extract 200mg/kg and MMC 0.1mg/kg is improved over 30%. 3. When Jukyeopseokgotanggagambang extract and MMC are administerated together, the weight of tumor is more decreased than MMC alone. 4. The effects of the Jukyeopseokgotanggagambang extract and MMC on the lysosomal enzymes in Ehrlich ascites carcinoma cell are more significantly improved than MMC alone. 5. Jukyeopseokgotanggagambang extract also increased the uptake of MMC into Ehrlich carcinoma cells. According to the above results, it could be suggested that Jukyeopseokgotanggagambang extract has indirect autitumor effects by strengthening the effects of MMC on tumor cells.

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죽엽석고탕가감방(竹葉石膏湯加減方)이 항암화학요법제(抗癌化學療法劑)의 세포독성(細胞毒性)과 종양세포(腫瘍細胞)의 라이소솜에 미치는 영향(影響) (Effects of Jukyeopseokgotanggagambang on Anti-tumor Chemotherapeutic Cytotoxicity and Lysosomal Enzymes of Tumor Cell)

  • 전승훈;전병훈;원진희;문구;문석재
    • 대한한방종양학회지
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    • 제3권1호
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    • pp.149-167
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    • 1997
  • In order to investingate the effects of Jukyeopseokgotanggagambang Extract on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI 1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficiency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. According to the change of colony-forming efficiency and SRB assay of Caki-1 cell, hep3B and A549 cells after exposure to the extract of Jukyeopseokgotanggagambang extract, that extract depressed the growth of tumor cells depending on its concentration. 2. Antitumor activities of the ethanol extract from Jukyeopseokgotanggagambang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of Jukyeopseokgotang-gagambang extract 200mg/kg and MMC 0.1mg/kg is improved over 30%. 3. When Jukyeopseokgotanggagambang extract and MMC are administerated together, the weight of tumor is more decreased than MMC alone. 4. The effects of the Jukyeopseokgotanggagambang extract and MMC on the lysosomal enzymes in Ehrlich ascites carcinoma cell are more significantly improved than MMC alone. 5. Jukyeopseokgotanggagambang extract also increased the uptake of MMC into Ehrlich carcinoma cells. According to the above results. it could be suggested that Jukyeopseokgotanggagambang extract has indirect autitumor effects by strengthening the effects of MMC on tumor cells.

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소요산가미방(逍遙散加味方)이 항암화학요법제(抗癌化學療法劑)의 항종양효과(抗腫瘍效果) 및 종양세포(腫瘍細胞)의 lysosomal enzymes에 미치는 영향(影響) (Effects of Soyosangamibang on antitumor chemotherapy and lysosomal enzymes of tumor cel)

  • 조현주;원봉희;문구;문석재;전병훈
    • 대한한의학회지
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    • 제18권2호
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    • pp.119-136
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    • 1997
  • In order to investigate the effects of Soyosangamibang Extract(逍遙散加味方抽出液) on antitumor effects after human cell lines(A549, hep3B, Caki-1, Ehrlich) transplantation into the peritoneal cavity or right groin in mice induced by RPMI1640 and GIBCO etc., the extracts of its herbal medicines were orally administered for 10 or 12 days. Experimental studies were performed for measurance of antitumor effect of MMC(Mitomycin C) and lysosomal enzyme's activities using colony forming efficency, SRB assay which were regarded as a valuable method for antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows: 1. The change of colony-forming efficiency and SRB assay of Caki-1 cells, hep3B and A549 cells after exposure to the extract of Soyosangamibang extract depressed the growth of tumor cells by concentration of Soyosangamibang, 2. Antitumor activity of the ethanol extract from Soyosangamibang extract and MMC on ascites form of Ehrlich carcinoma in mice is a little improved. Especially mean survival times of the group of 200mg/kg and MMC 0.1mg/kg is improved Over 50%. 3. WhenSoyosangamibang extract and MMC are administrated together, the weight of turnor is more decreased than MMC alone. 4. The lysosomal enzyme's activities of the Soyosangarmibang extract and MMC are more significantly improved than MMC alone. According to the above results, it could be suggested that Soyosangamibang extract has indirect antitumor effect by strengthen the effect of MMC.

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소적보중환(消積保中丸)의 항종양효과(抗腫瘍效果)에 대한 실험적(實驗的) 연구(硏究)

  • 노훈정;전병훈;문구;문석재
    • 대한한방종양학회지
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    • 제2권1호
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    • pp.43-56
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    • 1996
  • The sprig of Sojekbojungwhan(消積保中丸) has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Sojekbojungwhan extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell lines(A549, hep3B, Caki-1, Sarcoma 180) after esposure to Sojekbojungwhan extract using in ILS, colony forming efficiency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Sojekbojungwhan extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Sojekbojungwhan extact showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Sojekbojungwhan decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Sojekbojungwhan also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Sojekbojungwhan extract into TBM. 4. Sojekbojungwhan extract also increased NK cell activities. According to the above results, it could be suggested that Sojekbojungwhan extract has some antitumor effects.

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두 개의 다른 마우스 배아줄기세포의 차별적인 조혈세포 분화능 (Different Potential of Hematopoietic Differentiation in Two Distinct Mouse Embryonic Stem Cells)

  • 김진숙;강호범;송지연;오구택;남기환;이영희
    • 한국발생생물학회지:발생과생식
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    • 제9권2호
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    • pp.105-114
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    • 2005
  • 배아 줄기세포는 세포 치료 목적을 위한 재료로써 매우 큰 잠재력을 가지고 있으며, 이러한 잠재력의 실현을 위해서 세포의 운명에 결정적인 역할을 하는 요소들을 확인하고 특정 세포의 대량 생산을 위한 방법을 개발하여야 한다. 조혈과정은 폭넓게 연구되어 왔으며, 배아 줄기세포로부터 조혈세포의 분화는 lineage commitment에 관한 연구에 좋은 모델이 된다. 본 연구에서는, 두 종류의 마우스 배아 줄기세포주 TC-1과 B6-1를 이용하여 그 특성과 조혈세포 분화능을 비교하여 보았다. 두 세포주는 작은 차이는 있으나 줄기세포로서의 특성을 공통적으로 가지고 있었다. 그러나 methylcellulose 배양 system을 사용하여 embryonic body 형성능을 확인한 결과 TC-1이 B6-1에 비해 월등함을 확인하였다. 조혈세포 분화의 추적을 위해 blast colony의 형성, progenitor assay, RT-PCR을 통한 조혈세포 분화 관련 marker의 발현 분석을 수행한 결과, TC-1은 정상적으로 조혈세포를 생성해 내지만, B6-1은 제대로 분화되지 못함을 확인할 수 있었다. 이러한 결과들은 in vitro에서 배아 줄기세포로부터 조혈세포로 분화를 유도할 때, 보다 적합한 세포주의 탐색이 요구됨을 제시하며 이는 향후 인간 배아 줄기세포주에서도 마찬가지로 적용될 수 있음을 암시한다고 사료된다.

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