Related Experiment Video
Updated: May 3, 2026
Establishment of Orthotopic Patient-derived Xenograft Models for Brain Tumors using a Stereotaxic Device
Published on: May 2, 2025
A patient tumor-derived orthotopic xenograft mouse model replicating the group 3 supratentorial primitive
Zhigang Liu1, Xiumei Zhao1, Yue Wang1
1Diana Helis Henry Medical Research Foundation, New Orleans, Louisiana (Z.L., X.N.L); Laboratory of Molecular Neuro-Oncology, Texas Children's Cancer Center, Houston, Texas (Z.L., X.Z., Y.W., H.M., M.K., L.Q., X.N.L.); Texas Children's Cancer Center, Houston, Texas (P.A.B., T.K.M., J.M.S., L.P., C.C.L., M.C.); Department of Pathology, Texas Children's Hospital, Baylor College of Medicine, Houston, Texas (A.A.); Division of Hematology-Oncology, Arthur and Sonia Labatt Brain Tumor Research Center, Department of Pediatrics, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada (D.P., K.C. H., A.H.).
Background:
Supratentorial primitive neuroectodermal tumor (sPNET) is a malignant brain tumor with poor prognosis. New model systems that replicate sPNET's molecular subtype(s) and maintain cancer stem cell (CSC) pool are needed.
Methods:
A fresh surgical specimen of a pediatric sPNET was directly injected into the right cerebrum of Rag2/SCID mice. The xenograft tumors were serially sub-transplanted in mouse brains, characterized histopathologically, and subclassified into molecular subtype through qRT-PCR and immunohistochemical analysis. CSCs were identified through flow cytometric profiling of putative CSC markers (CD133, CD15, CD24, CD44, and CD117), functional examination of neurosphere forming efficiency in vitro, and tumor formation capacity in vivo. To establish a neurosphere line, neurospheres were propagated in serum-free medium.
Results:
Formation of intracerebral xenograft tumors was confirmed in 4 of the 5 mice injected with the patient tumor. These xenograft tumors were sub-transplanted in vivo 5 times. They replicated the histopathological features of the original patient tumor and expressed the molecular markers (TWIST1 and FOXJ1) of group 3 sPNET. CD133(+) and CD15(+) cells were found to have strong neurosphere-forming efficiency in vitro and potent tumor-forming capacity (with as few as 100 cells) in vivo. A neurosphere line BXD-2664PNET-NS was established that preserved stem cell features and expressed group 3 markers.
Conclusion:
We have established a group 3 sPNET xenograft mouse model (IC-2664PNET) with matching neurosphere line (BXD-2664PNET-NS) and identified CD133(+) and CD15(+) cells as the major CSC subpopulations. This novel model system should facilitate biological studies and preclinical drug screenings for childhood sPNET.
