Establishment of a Novel IDH1 wild type Glioblastoma Cell Line, SHG142, and Its Organoids

Glioblastoma (GBM) is the most aggressive primary malignant brain tumor and is associated with limited treatment options and poor prognosis. Conventional GBM cell lines undergo genetic drift and progressive divergence from patient-relevant molecular features over long-term culture, limiting their translational relevance. This study aimed to establish and characterize a novel isocitrate dehydrogenase 1 (IDH1)-wildtype GBM cell line, SHG142, along with its associated glioma stem-like cells (GSCs) and organoid models, to advance GBM biology and therapeutic research.

Primary tumor tissue from a 70-year-old female patient was cultured under standard conditions to generate the SHG142 cell line, which was authenticated by short tandem repeat (STR) profiling. Phenotypic and genetic features were evaluated using immunofluorescence (IF), karyotyping, and whole-exome sequencing (WES) including a concordance analysis between an early bulk primary culture (P2) and late-passage SHG142 (P50). GSCs were isolated via serum-free culture and magnetic-activated cell sorting (MACS). Tumorigenicity and invasiveness were assessed through intracranial xenografts in nude mice (n = 6/group) and coculture with brain organoids derived from human embryonic stem cells (n = 4 independent organoids).

SHG142 cells exhibited stable morphology and proliferation over more than 50 passages, retained key GBM markers (e.g., Nestin, Glial Fibrillary Acidic Protein (GFAP), O6-methylguanine-DNA methyltransferase (MGMT), Ki-67, P53, S-100, CD34), and exhibited chromosomal instability (+7, -10, and t(9;13)). WES revealed mutations in the TERT promoter, TP53 and NF1 and supported lineage continuity from P2 to P50 through shared SNVs and conserved driver events. CD133+ SHG142 GSCs demonstrated stem-like properties and invasive behavior in vivo and in organoid coculture. Compared with xenografts derived from SHG142 cells, GSC-derived xenografts were associated with increased malignancy and shorter survival (median survival 25 vs. 31.5 days, p = 0.025). The organoid coculture model confirmed deep infiltration by proliferative, stem-like tumor cells.

SHG142 and its GSCs represent a phenotypically stable, genetically defined GBM model with evidence of lineage continuity and retention of key driver events during in vitro propagation. Their tumorigenic and invasive properties make them valuable tools for mechanistic studies and preclinical therapeutic screening, supported by initial two-dimensional and three-dimensional drug response readouts that enable first-pass evaluation of therapeutic response.