Drug Assays on Cultures of Human Tumour Biopsies

Ambrose, E. J.; Andrews, Russel D.; Easty, DorothyM.; Field, E. O.; Wylie, J. A. H.

doi:10.1016/s0140-6736(62)92645-4

Cited by 22 publications

(5 citation statements)

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“…If more information were available about the tumour's sensitivity to the cytotoxic agents available, many more profitable perfusions might be seen. The effect of drugs on tumour cells in vitro (Ambrose et al 1962, Cobb et al 1961, Krementz et al 1959, Wright et al 1957, tumour implants into the anterior chamber of guinea-pigs (Creech 1961, personal communication), and enzyme studies (Wolberg & Curreri 1961) have all been tried, and their further evaluation is awaited.…”

Section: Discussionmentioning

confidence: 99%

Extracorporeal Arterial Perfusion

1963

Proceedings of the Royal Society of Medicine

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Section: Discussionmentioning

confidence: 99%

Extracorporeal Arterial Perfusion

1963

Proceedings of the Royal Society of Medicine

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“…quantities into Ambrose tubes (Ambrose et al, 1962) and incubated at 370 C. in a gas phase of 5% CO2 in air. A good monolayer of cells on the base of the tubes has usually formed in 48 hours and the medium is then changed, giving a clear fluid through which observations with an inverted microscope can be made.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The monolayer is almost always dense enough to allow counting to be performed with the Chalkley eyepiece (Curtis, 1960). The drugs selected for administration are then added according to the method already described (Ambrose et al, 1962), the same volume-0.08 ml.-of Hanks's saline being added to the control tubes.The drugs chosen for this series were: melphalan (p-di-(2-chloroethyl)amino-L-phenylalanine), thiotepa (triethylene thiophosphoramide), chlorambucil (p-di-(2chloroethyl)aminophenylbutyric acid), mannitol busul-F …”

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confidence: 99%

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Screening of 12 Gliomata Against Chemotherapeutic Agents in Vitro

Easty¹,

Wylie²

1963

BMJ

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Using the methods described in detail below, we have attempted to cultivate many types of tumours, and whereas we have had scant success with most carcinomata and sarcomata, our experience with gliomata, comprising astrocytomata, ependymomata, and medulloblastomata, has been almost invariably successful. The results obtained in the testing of 12 gliomata and a culture of HeLa cells are reported here. Methods and MaterialsThe specimens examined were all obtained from Atkinson Morley's Hospital or the Hospital for Sick Children at Great Ormond Street. At operation the, tumours are placed in either a dry sterile screw-capped bottle or into Parker's TC medium 199 and packed in ice in a vacuum flask. They are brought to the Chester Beatty Research Institute and arrive within the hour. Immediately on arrival the specimens are chopped up manually with fine scissors and, being very soft, fragment easily. The "mush" so obtained is transferred to a 0.06% solution of commercial trypsin (" difco ") in Parker's TC medium 199 and agitated in a magnetic stirrer for 30 minutes at 370 C. Thereafter the coarser particles in the suspension are allowed to settle and the supernatant fluid is pipetted off and centrifuged for five minutes at 1,500 r.p.m. The supernatant from this procedure, which contains the trypsin, is removed and the centrifuged cells are resuspended in the final medium, which consists of 20% of the patient's serum +80% Evans N.C.T.C. medium 109, to which folic acid (0.4 mg. /litre) and insulin (438 mg. /litre) have been added. Repeated washings of the cells were found to give less satisfactory results, and it is felt that the trypsin inhibitors in the serum incorporated in the medium are sufficient to neutralize the traces of trypsin remaining after centrifugation.The suspension thus achieved is dispensed in 0.3-ml. quantities into Ambrose tubes (Ambrose et al., 1962) and incubated at 370 C. in a gas phase of 5% CO2 in air. A good monolayer of cells on the base of the tubes has usually formed in 48 hours and the medium is then changed, giving a clear fluid through which observations with an inverted microscope can be made. The monolayer is almost always dense enough to allow counting to be performed with the Chalkley eyepiece (Curtis, 1960). The drugs selected for administration are then added according to the method already described (Ambrose et al., 1962), the same volume-0.08 ml.-of Hanks's saline being added to the control tubes.The drugs chosen for this series were: melphalan (p-di-(2-chloroethyl)amino-L-phenylalanine), thiotepa (triethylene thiophosphoramide), chlorambucil (p-di-(2-chloroethyl)aminophenylbutyric acid), mannitol busul-

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“…med. J., 1964, 2, 490-491 In previous communications (Ambrose, Andrews, Easty, Field, and Wylie, 1962 ;Easty and Wylie, 1963) a method was described for the preparation of monolayer tissue cultures from human biopsy specimens. The method was applied to the quantitative analysis of the effect of chemotherapeutic agents on human tumour specimens.…”

Section: Preliminary Communicationsmentioning

confidence: 99%