Development of a protocol to examine red blood cell functional adaptations to intermittent hypoxia

Abstract

This thesis presents the development of a standardized protocol for investigating the effects of intermittent hypoxia (IH) on human red blood cell (RBC) physiology. While IH contributes to conditions such as sleep apnea and cardiovascular disease, its direct impact on RBC function remains poorly characterized. The protocol isolates IH exposure as an independent variable and integrates in vitro stress testing (glucose deprivation, oxidative stress, hyperthermia) to evaluate RBC vulnerability, demonstrated by phosphatidylserine externalization, intracellular calcium accumulation, and reactive oxygen species generation indicating premature damage and cell death. Methodological considerations emphasize non-invasive blood sampling, minimal-risk hypoxic exposure, and reproducible lab methods to ensure feasibility and broad applicability. Rather than experimental finings, this work delivers a rigorously designed and justified framework that enables future research to assess IH-induced RBC dysfunction. The resulting protocol fills a current methodological gap and provides a reproducible foundation for studies in both pathological and high-altitude contexts.