Diffuse intraparenchymal gliomas (DIPGs) originate ventral to the pontine brain and develop over a certain period of time in children, suggesting that the disease may be associated with neural stem cells that can differentiate into neuronal or glial cells. The presence of neural stem cells under the ventricular canal of the lateral ventricles, which are also “tumor stem cells,” suggests that the disease may be associated with neural stem cells that can differentiate into neuronal or glial cells. Similar neural stem cells may also be present around the third and fourth ventricles, which are involved in DIPGs generation. In this study, we established cell lines using pontine glioma specimens obtained by autopsy and found that neural stem cells in the ventral part of the pontine brain may be the source of this tumor and that the Hedgehog signaling pathway is involved in the occurrence and development of DIPGs. 1. Distribution of neural stem cells in the brainstem It was found that two different types of nestin-positive cells exist in the postnatal human brainstem: located in the ventricles of the four ventricles. subcanalicular and ventral to the pontine brain. The neural stem cells located in the pontocerebral retest had elongated protrusions with a bipolar distribution, as well as positive for the neural stem cell marker protein vimentin (Vimentin), but did not express glial acidic protein (GFAP). Nearly half of the cells were Olig2-positive. In infancy, these cells were predominantly distributed ventral to the brainstem, significantly diminished at 2 years of age, and the distribution of this cell increased significantly ventral to the pontine brain at 5-7 years of age, peaking at 6 years of age. The second peak of ventral nestin-positive cells in the pontocerebrum coincided with the age of prevalence of DPIGs. 2. Hh pathway activation and neural stem cell differentiation correlate The subventricular neural stem cells in the four ventricles of mice are similar to those in humans, but ventral pontocerebral nestin and waveform filament protein positive cells are rare, but Olig2-positive cells are more distributed, and these cells show other characteristics of primitive cells. The differentiation of these stem cells can be promoted by activating the Hh pathway. These stem cells begin to appear ventral to the brainstem 14 days after birth in animals and gradually increase, corresponding to three times the number at 14 days by 21 days, a time period that corresponds to 5-7 years of age in children by developmental stage. Observation of medulloblastoma animals with Hh pathway activation revealed a significantly earlier increase in neural stem cells ventral to the brainstem. The number of neural stem cells ventral to the brainstem was significantly reduced 21 days after birth. It shows that Hh pathway can promote the development of neural stem cells. 3. Correlation between diffuse glioma cells in the ventral part of the pontine brain and neural stem cells DIPGs tumors (WHO III, mesenchymal astrocytoma) with high expression of nestin and GFAP were used in 5-year-old patients. By culturing these tumor cells, tumor neurospheres can be formed, similar to neurosphere formation by neural stem cells. The decidualized tumor cells can form neurospheres again (secondary neurospheres). When these tumor cells were injected into the lateral and quadrigeminal ventricles of nude mice, the tumors showed invasive growth. In animals injected into the lateral ventricles, tumors grew around the ventricles, the brain and even the cerebellum; in animals injected into the four ventricles, tumors invaded the cerebellum and brainstem, resembling the manifestation of invasive brainstem tumors. 4. the role of Hh pathway in DIPGs In the study, the expression of Hh pathway receptor system in brainstem tumors was found. Hh pathway antagonists significantly decreased the formation of secondary neurospheres in tumor cells (67% decrease, p<0.001); increasing the activity of Hh pathway, the formation of secondary neurospheres increased significantly (200%, p<0.05). The present study shows that DIPGs may originate from neural stem cells in the ventral part of the brainstem, and the Hh pathway system that promotes stem cell development may be involved in the occurrence and development of DIPGs.