• Title/Summary/Keyword: Colony forming efficiency assay

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

  • Jo, Eunhye;Lee, Jaewoo;Park, Sun-Young;Kim, Pilje;Choi, Kyunghee;Eom, Igchun
    • Journal of Environmental Health Sciences
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    • v.41 no.1
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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.

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 (자기 활성 세포 분리법과 군체 분리법으로 분리된 건 줄기세포의 자가 재생 능력 및 분화능 효율 비교)

  • Lee, Moses;Choi, Yoorim;Yoon, Dong Suk;Lee, Jin Woo;Yoon, Gil Sung;Choi, Woo Jin;Han, Seung Hwan
    • Journal of Korean Foot and Ankle Society
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    • v.18 no.3
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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

  • No, Hun-Jeong;Jeon, Byeong-Hun;Mun, Gu;Mun, Seok-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.2 no.1
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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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Experimental Studies on the Change of Cytotoxic and Antitumor Effects according to the Prebrewed Method of Semen Tiglii and Rhizoma Coptidis (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))

  • Jo, Soung-Gak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.1 no.1
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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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Influence of Gamihagochosan on The Antitumor effect of Anticancer Drug and The Proliferation of Tumor Cell Lines (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))

  • Kim, Gyun-Taek;Jeon, Byeong-Hun
    • The Journal of Internal Korean Medicine
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    • v.18 no.1
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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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Effects of Jukyeopseokgotanggagambang on Anti-tumor Chemotherapeutic Cytotoxicity and Lysosomal Enzymes of Tumor Cell (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)

  • Jeon, Seoung-Hun;Moon, Gu;Jeon, Byung-Hoon
    • The Journal of Internal Korean Medicine
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    • v.18 no.1
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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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Effects of Jukyeopseokgotanggagambang on Anti-tumor Chemotherapeutic Cytotoxicity and Lysosomal Enzymes of Tumor Cell (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))

  • Jeon, Seung-Hun;Jeon, Byeong-Hun;Won, Jin-Hui;Mun, Gu;Mun, Seok-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.3 no.1
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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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Effects of Soyosangamibang on antitumor chemotherapy and lysosomal enzymes of tumor cel (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))

  • Cho, Hyun-Ju;Won, Bong-Hui;Mun, Gu;Mun, Seok-Jae;Jeon, Byeong-Hun
    • The Journal of Korean Medicine
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    • v.18 no.2
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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)

  • No, Hun-Jeong;Jeon, Byeong-Hun;Mun, Gu;Mun, Seok-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.2 no.1
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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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Different Potential of Hematopoietic Differentiation in Two Distinct Mouse Embryonic Stem Cells (두 개의 다른 마우스 배아줄기세포의 차별적인 조혈세포 분화능)

  • Kim, Jin-Sook;Kang, Ho-Bum;Song, Jee-Yeon;Oh, Goo-Taeg;Nam, Ki-Hoan;Lee, Young-Hee
    • Development and Reproduction
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    • v.9 no.2
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    • pp.105-114
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    • 2005
  • Embryonic stem(ES) cells have tremendous potential as a cell source for cell-based therapies. Realization of that potential will depend on our ability to understand and manipulate the factors that influence cell fate decision and to develop methods for getting enough cell numbers for clinical applications. Hematopoiesis has been widely studied, and hematopoietic differentiation from ES cells is a good model to study lineage commitment. In this study, we investigated stemness and compared the efficiency of hematopoietic differentiation using two different mouse embryonic stem cell lines TC-1 and B6-1. Although the two cell lines showed known stem cell properties with minor differences, the embryoid body formation efficiency in methylcellulose was much higher in TC-1 than B6-1. When measured potentials of hematopoietic differentiation using functional(colony-forming cell) and phenotypic(specific marker expression) assays, we found that TC-1 can differentiate into hematopoietic cells in methylcellulose culture but B6-1 cannot. These results imply that we can improve the efficiency of hematopoietic cell differentiation by selection of proper cell lines and this may be also applied in the differentiation of human embryonic stem cells.

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