Establishment of the glioma polyploid giant cancer cell model by a modified PHA-DMSO-PEG fusion method following dual drug-fluorescence screening in vitro.

Journal of neuroscience methods(2021)

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摘要
BACKGROUND:In glioma, cell fusion and the number of the polyploid giant cancer cells (PGCC) were found to be augmented with tumor grades (WHO Ⅰ-Ⅳ) and closely related to poor prognosis. However, the pathological and molecular characteristics of glioma PGCCs remain unclear due to the lack of cell model in vitro and in vivo. NEW METHOD:Here, we reported a novel approach to obtain the glioma PGCCs by the PHA-DMSO-PEG fusion method following dual drug-fluorescence screening in vitro. Glioma cells were labelled by lentiviruses infection and fusion hybrids were obtained by puromycin screening and fluorescence-activated cell sorting (FACS). RESULTS:Glioma tumor-tumor cell fusion efficiency was significantly improved by PHA and DMSO. Glioma PGCCs were successfully obtained after puromycin screening and FACS. Cell size, DNA content and chromosome numbers of the glioma PGCCs were almost twice than that of the parental glioma cells. Moreover, glioma PGCCs showed a decreased proliferation rate but enhanced temozolomide resistance potential compared to the parental cells. COMPARISON WITH EXISTING METHODS:We firstly obtained the glioma PGCCs by a modified fusion method in vitro. The fusion efficiency of the PHA-DMSO-PEG fusion method was much higher compared to PEG fusion method. Moreover, the dual drug-fluorescence screening method was more convenient and effective compared to the single one. CONCLUSIONS:We successfully established the glioma PGCC model through a modified PHA-DMSO-PEG fusion method following dual drug-fluorescence screening in vitro. Glioma PGCCs showed a deceased proliferation rate but increased TMZ resistance capacity.
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