Sleep Disorders: Sleep Disorders III |

Development of Cell Culture Model of Intermittent Hypoxia FREE TO VIEW

Xiaohong Cai; Hongfang Mei; Hongxia Wang; Fangfang Hong; Liya Chen; Jing Lin
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Department of Pulmonolory, Second Affiliated Hospital & Yuying Children′s Hospital, Wenzhou Medical University, Wenzhou, China

Copyright 2016, American College of Chest Physicians. All Rights Reserved.

Chest. 2016;149(4_S):A579. doi:10.1016/j.chest.2016.02.604
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SESSION TITLE: Sleep Disorders III

SESSION TYPE: Original Investigation Poster

PRESENTED ON: Saturday, April 16, 2016 at 11:45 AM - 12:45 PM

PURPOSE: To establish and validate a novel model of cultured cells for imitating intermittent hypoxia.

METHODS: In a chamber with experiment cabin and simulated air control cabin, we decided the frequency and duration of the intermittent hypoxia model according to the time of hypoxia and reoxygenation. The subcultured A549 cells were randomly divided into 7 groups, respectively named as control group (C); 6 h intermittent hypoxia group (6IH); 9 h intermittent hypoxia group (9IH); 6 h simulated air control group (6AC); 9 h simulated air control group (9AC); 4 h sustained hypoxia group (4SH); 6 h sustained hypoxia group (6SH). When the model establishment finished, the cellular morphology was observed under inverted microscope. The mRNA expression of HIF-1α was detected by real-time fluorescence quantitative PCR. The protein expression of HIF-1α was determined by immunohistochemistry.

RESULTS: The intermittent hypoxia cycle was 5% O2 60 min/20% O2 30 min for 6 cycles. The damaged A549 cells was observed in 6IH group, 9IH group and 6SH group among which the 9IH group was the most serious one. Compared with 6IH group, the expression of HIF-1α at mRNA and protein levels was significantly increased in 9IH group (P<0.05). The expression of HIF-1α at mRNA and protein levels in 6IH group and 9IH group was higher than that in 4SH group and 6SH group respectively (P<0.05). No significant difference among the control group, 6AC group and 9AC group was found (P>0.05).

CONCLUSIONS: The model of 5% O2 60 min/20% O2 30 min for 6 cycles can simulate the pathological mechanism of OSAHS. This model is suitable for studying intermittent hypoxia in adherent cells.

CLINICAL IMPLICATIONS: To establish and validate a novel model of cultured cells for imitating intermittent hypoxia.

DISCLOSURE: The following authors have nothing to disclose: Xiaohong Cai, Hongfang Mei, Hongxia Wang, Fangfang Hong, Liya Chen, Jing Lin

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