A general native-state method for determination of proliferation capacity of human normal and tumor tissues in vitro.

Hoffman, Robert M.; Connors, Kenneth M.; Meerson-Monosov, Ann Z.; Herrera, Hector; Price, Jeffrey H.

doi:10.1073/pnas.86.6.2013

Cited by 49 publications

(18 citation statements)

References 17 publications

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“…The brightgreen nuclear staining is due to silver grains being exposed due to [3H]thymidine incorporation. The polarized light is then reflected as bright green (30). Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The slides were then analyzed with a polarizing microscope at x 400 power in a blinded fashion (30). Replicating cells were easily determined by the presence of bright-green-reflecting silver grains over the cell nuclei.…”

Section: Methodsmentioning

confidence: 99%

“…Thus we have developed a native-state technique of histoculturing tumors in three dimensions on flexible collagencontaining gels with the maintenance of native tumor histology and function and with a high success rate of sustained growth in culture (28)(29)(30). In the native-state system, longterm growth can be achieved, which can in some instances be Proc.…”

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confidence: 99%

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Cancer biology for individualized therapy: correlation of growth fraction index in native-state histoculture with tumor grade and stage.

Vescio

Connors²,

Youngkin³

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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“…The brightgreen nuclear staining is due to silver grains being exposed due to [3H]thymidine incorporation. The polarized light is then reflected as bright green (30). Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Cancer biology for individualized therapy: correlation of growth fraction index in native-state histoculture with tumor grade and stage.

Vescio

Connors²,

Youngkin³

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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“…26) A histoculture method was developed by modifying a tissue culture method to make it three-dimensional with similar use of ECM, and employed for long-term culture of primary tumor cells. [27][28][29] Using this culture method, Hoffman et al 30) and Furukawa et al established the histoculture drug response assay (HDRA), and have reported its clinical utility. [31][32][33] However, in their method, the concentration of the applied drug was tens or hundreds of times higher when converted to AUC, whereas in CD-DST, the concentration of the applied drug is close to the in vivo level, which appears to be more reasonable.…”

Section: Discussionmentioning

confidence: 99%

Examination of in vitro Chemosensitivity Test Using Collagen Gel Droplet Culture Method with Colorimetric Endpoint Quantification

Kobayashi¹,

Higashiyama

Minamigawa³

et al. 2001

Japanese Journal of Cancer Research

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To develop a simpler method of performing the collagen gel droplet-embedded culture drug sensitivity test (CD-DST), we examined the introduction of colorimetric quantitative determination of images for evaluation of anticancer effect against cancer cells alone in the presence of fibroblasts, based on differences in proliferative morphology and stainability with neutral red of cells within collagen gel drops determined using a video-microscope and NIH Image software. In examinations using a human cancer cell line and a fibroblast cell line, a high degree of linearity between number of cancer cells and image-optical density was found within the range of 10 2 -10 6 cells/droplet (r 2 = = = =0.933). Using NIH Image, fibroblast cells could be eliminated at a cut-off value of 128, and an immunocytochemical method demonstrated that the cells eliminated from the image were indeed fibroblasts, and those remaining were cancer cells. CD-DST was carried out with mixtures of cancer cells with fibroblasts at various ratios, and the feasibility of evaluating anticancer activity in cancer cells alone with no effect of fibroblasts at any mixing ratio was confirmed. In addition, for CD-DST of primary cell cultures of human lung cancers collected at the time of surgery, a high correlation between results obtained with the volume supplementation method, a current cell quantification method, and those with the imaging colorimetric quantification method was obtained (r = = = =0.933). These results indicate that introduction of imaging colorimetric quantification utilizing NIH Image makes CD-DST a quick and simple method that should be highly useful for clinical chemosensitivity testing using primary cell cultures of human cancers.

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“…24,25 Briefly, each tumor specimen was minced finely and aseptically with a scalpel, which was then placed on each of the prepared collagen surfaces in 24-well plates. The plates were incubated at 37 C for 7 days with the drug dissolved in RPMI 1640 medium containing 20% fetal calf serum in a…”

Section: Quantification Of Apoptotic Cells In Tissue Sectionsmentioning

confidence: 99%

Combination of p53 codon 72 polymorphism and inactive p53 mutation predicts chemosensitivity to 5‐fluorouracil in colorectal cancer

Tominaga

Itō

Takifuji

et al. 2010

Intl Journal of Cancer

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There are increasing reports showing the clinical significance of the p53 polymorphism status in terms of the response to chemotherapy. We investigated whether p53 polymorphism and mutation were associated with in vitro sensitivity to 5-fluorouracil (5-FU) in patients with colorectal cancer. Chemosensitivity to 5-FU was evaluated by the collagen gel droplet embedded culture drug sensitivity test. 5-FU sensitivity of tumor cells without inactive p53 mutation in the arginine/arginine (Arg/Arg) variant was significantly higher than that of tumor cells with or without inactive p53 mutation in other variants (p 5 0.022), whereas the 5-FU sensitivity of tumor cells with inactive p53 mutation in the Arg/Arg variant was significantly lower than that of tumor cells with or without inactive p53 mutation in other variants (p 5 0.002). In the Arg/Arg variant, apoptotic cells induced by 5-FU treatment in patients without inactive p53 mutation were more markedly increased than those in patients with inactive p53 mutation (p 5 0.037). Bax and Bcl-2 protein expressions in tumor tissue treated with 5-FU were associated with both 5-FU sensitivity and the apoptotic cell count. Our data show that the Arg/Arg genotype without inactive p53 mutation could be predictive of a more favorable response and the Arg/Arg genotype with inactive p53 mutation a less favorable response to chemotherapy using 5-FU in CRC. The combination of the p53 codon 72 polymorphism and p53 mutation status is a potential predictive marker of sensitivity to 5-FU in CRC.The p53 tumor suppressor gene encodes a nuclear protein that induces growth arrest or apoptosis in response to cellular stress. 1 Apoptosis is a fundamental mechanism by which DNA-damaging anticancer agents cause cytotoxicity. 2 p53 has a major function in transducing stress to the apoptotic machinery of cells, consistent with the importance of the p53 status as a determinant of the cellular response to DNA-damaging drugs. 3,4 The presence of an intact p53 function confers tumor sensitivity to DNA-damaging agents such as cisplatin. 5 However, p53 is mutated in at least 50% of human cancers. 6 The mutation site is widely distributed in DNA-binding domain including the amino acid part that is called hot spot as codons 175, 245, 248, 249, 273 and 282. In a recent analyses with a large number of patients, type of mutation (inactive TP53 mutations) and adjuvant treatment were identified as important factors in determining the prognostic significance of p53 mutation in colorectal cancer (CRC). 7,8 Another recent study has quantitated the functional activity of different p53 mutation with respect to their ability to transactivate target genes. There were 3 subtypes: mutants with no activity, mutants with significantly reduced but some residual activity and mutants with activity comparable with that of wild-type p53. 9 The most common p53 mutation in human tumors show a clear loss of transactivation activity. 10 We can designate p53 hot spot mutation as inactive p53 mutation. Inactive p53 mutation is impo...

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A general native-state method for determination of proliferation capacity of human normal and tumor tissues in vitro.

Cited by 49 publications

References 17 publications

Cancer biology for individualized therapy: correlation of growth fraction index in native-state histoculture with tumor grade and stage.

Cancer biology for individualized therapy: correlation of growth fraction index in native-state histoculture with tumor grade and stage.

Examination of in vitro Chemosensitivity Test Using Collagen Gel Droplet Culture Method with Colorimetric Endpoint Quantification

Combination of p53 codon 72 polymorphism and inactive p53 mutation predicts chemosensitivity to 5‐fluorouracil in colorectal cancer

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