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In vitro studies on the growth and chemosensitivity of short-term cultures derived from childhood brain tumours

Lewandowicz, Grazyna Maria; (1997) In vitro studies on the growth and chemosensitivity of short-term cultures derived from childhood brain tumours. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Clinical evidence indicates that many tumours of the central nervous system in children respond to radiotherapy. There is less evidence that these tumours are sensitive to chemotherapy. Unfortunately, there is a paucity of established cell lines from most types of paediatric brain tumour in which to investigate chemosensitivity in vitro. There have been no reports of cell lines derived from benign astrocytomas and only one derived from a malignant astrocytoma of childhood. Of the other tumour histologies, only two cell lines from ependymomas and a small number from medulloblastomas have been established. The aim of this study was to carry out a systematic examination of the conditions necessary to grow childhood brain tumours in vitro for chemosensitivity studies. Tumour biopsy material was enzymatically disaggregated with collagenase and plated onto substrates known to enhance the attachment and growth of human tumour cells, including Cell-tak, fibronectin, laminin, Matrigel and vitrogen. Marked differences were noted between the substrates in their ability to promote in vitro growth of these tumour cell lines. Cell lines grew best on plastic, fibronectin and laminin and reached passage levels in excess of 20. The phenotype of cells growing on a variety of substrates was examined. The intermediate filament composition of a number of cultures was determined using monoclonal antibodies against glial fibrillary acidic protein, neurofilament protein, vimentin, cytokeratin and desmin. Additionally, a polyclonal antibody against fibronectin and synaptophysin was used. Short-term cultures derived from tumours of glial origin expressed glial markers, for example astrocytomas expressed GFAP. Those cultures of neuronal origin (e.g., medulloblastomas) expressed neuronal markers such as neurofilament protein and synaptophysin. However, unexpected expression of certain markers was also observed, where a small number of astrocytoma-derived cultures expressed neurofilament protein (albeit to a small degree) and medulloblastoma-derived cultures occasionally expressed desmin. The chemosensitivity of these cultures was determined using the MTT assay. Neuroblastomas were extremely sensitive to vincristine, while ependymomas were markedly resistant. Astrocytomas and medulloblastomas were of similar sensitivity. Cultures derived from neuroblastoma were sensitive to CCNU, whilst the other tumour groups had similar sensitivities. There was little difference between the chemosensitivities to procarbazine but some medulloblastomas were very resistant to this drug. These results show similarities of chemosensitivity with adult malignant gliomas. This indicates the need for the identification of new drugs using in vitro screening based on short-term cultures derived from paediatric brain tumours.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: In vitro studies on the growth and chemosensitivity of short-term cultures derived from childhood brain tumours
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Health and environmental sciences; Chemosensitivity
URI: https://discovery.ucl.ac.uk/id/eprint/10105022
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