Abstract: Objective To develop a hypoxic experimental apparatus for nerve cells with good function and easy to use, which can be used to explore the pathogenesis and management of neonatal brain injury by simulating a hypoxic-ischemia process on a cellular level. Methods A set of hypoxic experimental apparatus was designed and developed. Using this hypoxic experimental apparatus, the oxygen-glucose deprivation (OGD) model of nerve cells were respectively established with various OGD time of 30 min, 45 min, 60 min, 2 h and 3 h, respectively. The influence of different OGD time on the survival rates of nerve cells was assessed 24 hours after OGD by using Cell Counting Kit-8 (CCK-8) and Hoechst33342/PI double fluorescence dying method. Results The set of hypoxic experimental apparatus consisted of four parts including a hypoxic chamber, a commercial source of non-oxygen gas, a water-sealed bottle for draining air, and a CO₂ incubator. The hypoxic chamber developed was lucid with good closure property and easiness to open and carry. The results for evaluating the influence of different OGD time on the survival rates of nerve cells established by using the hypoxic experimental apparatus showed that the apoptotic and necrotic cells tended to increase significantly with the extension of OGD period gradually. The survival rates of nerve cells were from 85% with 30min OGD reduced to 22% with 3 h OGD. About 45~60 min OGD could cause 40~55% of nerve cells to die. Conlusions The set hypoxic experimental apparatus developed is with good function and easy to use, which can ensure the successful establishment of OGD model of nerve cells.

Key words: Neonatal brain injury, Nerve cell, Oxygen-glucose deprivation, Cell hypoxic chamber