Hypoxic injury in developing glial cells and their differential response can result in neurological deficits in neonates. This study attempts to understand the role of autophagy in developing premyelinating oligodendrocytes (pre-OL), mature-OL and astrocytes exposed to hypoxic injury. Human fetal neural stem cell-derived pre-OL, mature OL and astrocytes were exposed to hypoxia (0.2% oxygen) and normoxia (20% oxygen) for 48 hours followed by RNA-sequencing. Transcriptomic analysis revealed upregulation of autophagy in mature OL and astrocytes but not in pre-OL. MO3.13 (resembling pre-OL) and SVG (astrocytes) cell lines were used for further experiments. MO3.13 cells were differentiated into mature OL using PMA, and characterized using specific markers (n=5). Pre-OL, mature OL and astrocytes were then subjected to hypoxia (0.2% oxygen) and normoxia (20% oxygen) for 48 hours. Hypoxia exposure was confirmed by increased expression of HIF-1α and CA9. Interestingly, severe hypoxia resulted in greater cellular oxidative stress only in pre-OL but not in mature OL or astrocytes, by the CellROX assay using flow cytometry. Analysis of mitochondrial membrane potential in severe hypoxia using JC-1 dye, revealed higher mitochondrial damage in pre-OL compared to mature OL and astrocytes (n=3). LC3B expression indicative of autophagic flux was found to increase in mature OL and astrocytes exposed to hypoxia as compared to pre-OL by Western blotting (n=3). These novel findings highlight that differential recruitment of autophagic defense response may be a mechanism underlining the vulnerability of pre-OL to hypoxic injury, and may provide potential therapeutic strategies in neonatal hypoxic brain injury.