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A novel three-dimensional “all human” in vitro brain tumor invasion model

Research output: Contribution to journalArticlepeer-review

  • S. Murray
  • H. Rooprai
  • R. Selway
  • Geoff Pilkington
In order to study brain tumor invasion in vitro, previous three-dimensional model systems have utilized pre-cultured chick heart fragment and rat brain as targets in co-culture with human brain tumor. We have now succeeded in establishing a novel three-dimensional model whereby human neoplastic glial cell spheroids are juxtaposed with spheroids derived from non-neoplastic human brain tissue. Spheroids are developed from either established glioma-derived cell lines or from primary cultures of glioma biopsy as well as brain resected from patients who have undergone surgery for epilepsy, short-term astrocyte-rich cell cultures derived therefrom. Using the “hanging drop” method, 45,000 cells were inversely suspended from a petri dish in 20 ml of DMEM. After either 24 h (fast-growing cells) or 48 h (slower growing cells) in their gravity pools, spheroids are transferred to agar-coated petri dishes for a further 24 h prior to confrontation. Cells were tracked with the use of fluorescent cell trackers; progress was recorded every 3 days for up to 15 days. Fluorescent cell trackers were also employed for time-lapse video microscopy in the most invasive combination over 5 days. We are further developing the model in utilizing human serum instead of fetal calf serum, which has been shown to alter antigenic expression and growth rate of human glioma cells in two-dimensional cultures within our laboratories. The effects of various agents that may impede local brain tumor invasion are currently under investigation. To these ends we are utilizing scanning and transmission electron microscopy, confocal microscopy, total internal reflected fluorescence (TIRF) microscopy, and live cell imaging utilizing Improvision Velocity and Openlab software on a Zeiss inverted axiovert 200M microscope. This work was supported by the Dr Hadwen Trust
Original languageEnglish
Pages (from-to)307-308
Number of pages2
JournalNeuro-Oncology
Volume7
Issue number3
DOIs
Publication statusPublished - 2005
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