Limitations of the agar colony-forming assay for the assessment of paediatric tumours

Ablett, G; Smith, Peter J.; Sheridan, J. W.; Lihou, Michelle G.

doi:10.1038/bjc.1984.213

Cited by 9 publications

(3 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Apart from being important in relation to understanding the actions of SPS and lysate, these findings emphasise the importance of selecting the correct time for scoring colonies Ablett et al (1984) have shown the need to consider the growth rates of primary human tumour-derived colonies when selecting the appropriate time to score such colonies. The decision as when to score would assume even greater importance if, in addition to the intrinsically slow growth rate of such colonies, cytotoxic regimens were to have an opposite effect on lag to that of SPS and REL in causing a prolongation of this phase.…”

Section: Clonogenic Cell Assaymentioning

confidence: 99%

Potentiation of anchorage-independent colony formation by sodium polyanethol sulphonate

Sheridan¹,

Bishop²,

Simmons³

et al. 1984

Br J Cancer

Self Cite

View full text Add to dashboard Cite

Section: Clonogenic Cell Assaymentioning

confidence: 99%

Potentiation of anchorage-independent colony formation by sodium polyanethol sulphonate

Sheridan¹,

Bishop²,

Simmons³

et al. 1984

Br J Cancer

Self Cite

View full text Add to dashboard Cite

“…Tumours samples were dissociated and cloned in agar as described by Ablett et al (1984). Primary cultures, were established in tissue culture flasks from dissociated tumour cells in RPMI 1640 with 10% FCS, were incubated at 37°C in an environment of 5% 02, 5% C02, 90% N2 and 100% humidity.…”

Section: Establishment Of Cell Culturesmentioning

confidence: 99%

Comparison of in vitro activity of epipodophyllotoxins with other chemotherapeutic agents in human medulloblastomas

Tomlinson

Lihou²,

Smith³

1991

Br J Cancer

View full text Add to dashboard Cite

Summary Surgical specimens from 15 medulloblastoma patients were used to establish early passage cultures. In vitro sensitivity to a battery of cytotoxic agents, including some in current medulloblastoma treatment protocols, was measured. Drug sensitivity was assessed at clinically relevant drug concentrations using the 3H-thymidine uptake method. Tumours were predicted to be sensitive if >37% were killed by exposure to drugs at clinically achievable levels. A poor response to vincristine (Vcr), cis-platin (CDDP), hydroxyurea (HU) Medulloblastoma is an important paediatric brain tumour because of its high incidence and malignant behaviour. The overall disease free survival at 5 years, in patients receiving surgery and craniospinal axis radiotherapy, is approximately 50%. This is considered to be the upper limit of curability by these treatment modalities. Chemotherapy has been shown to be advantageous in selected patient groups either as adjuvant therapy or as treatment of recurrent disease. The value of chemotherapy in the treatment of medulloblastoma will be dependent on the availability and identification of drugs active against this tumour. At present, protocols for chemotherapy usually include the lipophilic compounds carmustine (BCNU) or lomustine and procarbazine. Vincristine (Vcr), whilst poorly absorbed by cerebrospinal fluid and brain, is also commonly used (Bloom, 1986;Workman, 1986). Currently '8 in 1F therapy is being evaluated (Pendergrass et al., 1987).Cytotoxicity has been assessed by clonal assays in agar or on plastic, incorporation of 3H-thymidine into DNA, uptake of vital dyes and other methods with much the same results. Predictions of resistance are close to 100% correct; prediction of sensitivity, whilst less reliable, still has a success rate of around 70% Moon et al., 1981; Rosenbloom et al., 1983;Bogdahn, 1983;Kornblith et al., 1981;Kimmel et al., 1987). We have examined drug sensitivity in human medulloblastoma by using 14 early passage cultures. Our study provides data on the degree of heterogeneity of response between medulloblastomas obtained from a number of patients. Furthermore, the cells were from early passage cultures, and hence should give a better indication of chemosensitivity of the tumours in vivo. The epipodophyllotoxins teniposide (VM-26) and etoposide (VP16-213) were identified as potentially useful agents in this disease. We investigated the basis of primary clinical resistance to these drugs by quantitating intracellular steady-state drug concentrations, drug-induced protein-linked DNA strand breaks, and topoisomerase II expression and activity. Materials and methods Establishment of cell culturesAll studies were made on material collected at procedures performed for clinical reasons and remaining after sufficient tissue has been taken for clinical laboratory study. The histological diagnosis was confirmed by an independent neuropathologist (Professor B.W. Scheithauer). Over a period of 2 years, 16 medulloblastoma specimens were received from a total of 15 patients...

show abstract

“…The soft-agar clonogenic assay, which can measure the reproductive capacity of individual cells, is a reliable indicator of cell lethality after exposure to drugs and has a good clinical correlation (Von Hoff et al, 1981), but carries a number of technical disadvantages such as low plating efficiency and the long time required to obtain results. The poor growth of most softtissue sarcomas in this assay system also limits its application for this disease (Ablett et al, 1984). For drugs that directly or indirectly inhibit thymidylate biosynthesis, the in situ thymidylate synthase (TS) assay may be a useful in vitro test for predicting antifolate cytotoxicity (Yalowich et al, 1985).…”

mentioning

confidence: 99%

Prediction of antifolate efficacy in a rat sarcoma model

Lin

Chang

et al. 1991

Intl Journal of Cancer

View full text Add to dashboard Cite

A methylcholanthrene-induced rat sarcoma propagated both in vitro and in vivo was used to examine the usefulness of a rapid biochemical in situ assay that measures thymidylate synthase (TS) activity in whole cells to predict sensitivity and resistance to the folate antagonists methotrexate (MTX), 10-ethyl-10-deazaaminopterin (10-EDAM) and trimetrexate (TMTX). There was an excellent correlation between the effects of these drugs on inhibiting TS and cytotoxicity as measured by a clonogenic assay, when the exposure time was 4 hr and those when the rat sarcoma cell line was employed (p less than 0.001). When tumor-cell suspensions were prepared from the rat sarcoma propagated in vivo, they were less sensitive to these antifolates, but the relative effectiveness of the 3 antifolates was similar: TMTX much greater than 10-EDAM greater than MTX. As expected, continuous exposure (10 to 12 days) produced cytotoxicity at a much lower dose of these drugs. When the 3 drugs were tested for anti-tumor effectiveness in rats bearing this tumor, the tumor regression measured was best with TMTX; 10-EDAM was intermediate in effectiveness as compared with MTX. These results indicate that the in situ TS assay and clonogenic assay may be used to predict anti-tumor efficacy of antifolates in vivo in this rat sarcoma.

show abstract

Limitations of the agar colony-forming assay for the assessment of paediatric tumours

Cited by 9 publications

References 8 publications

Potentiation of anchorage-independent colony formation by sodium polyanethol sulphonate

Potentiation of anchorage-independent colony formation by sodium polyanethol sulphonate

Comparison of in vitro activity of epipodophyllotoxins with other chemotherapeutic agents in human medulloblastomas

Prediction of antifolate efficacy in a rat sarcoma model

Contact Info

Product

Resources

About