Circulation and cellular activity were restored in a pig’s brain four hours after its death which challenges long-held assumptions about the timing and irreversible nature of the cessation of some brain functions after death.
This may help doctors find ways to help salvage brain function in stroke patients, or test the efficacy of novel therapies targeting cellular recovery after injury.
Above -Immunofluorescent stains for neurons (green), astrocytes (red), and cell nuclei (blue) in a region of the hippocampus of a pig’s brain left untreated 10 hours after death (left) or subjected to perfusion with the BrainEx technology. Ten hours postmortem, neurons and astrocytes undergo cellular disintegration unless salvaged by the BrainEx system. (Image credit: Stefano G. Daniele & Zvonimir Vrselja; Sestan Laboratory; Yale School of Medicine)
It is unclear whether this approach can be applied to a recently deceased human brain. The chemical solution used lacks many of the components natively found in human blood, such as the immune system and other blood cells, which makes the experimental system significantly different from normal living conditions. However, the researcher stressed any future study involving human tissue or possible revival of global electrical activity in postmortem animal tissue should be done under strict ethical oversight.
The brain of a postmortem pig obtained from a meatpacking plant was isolated and circulated with a specially designed chemical solution. Many basic cellular functions, once thought to cease seconds or minutes after oxygen and blood flow cease, were observed, the scientists report.
Cellular death within the brain is usually considered to be a swift and irreversible process. Cut off from oxygen and a blood supply, the brain’s electrical activity and signs of awareness disappear within seconds, while energy stores are depleted within minutes. Current understanding maintains that a cascade of injury and death molecules are then activated leading to widespread, irreversible degeneration.
However, researchers in Sestan’s lab, whose research focuses on brain development and evolution, observed that the small tissue samples they worked with routinely showed signs of cellular viability, even when the tissue was harvested multiple hours postmortem. Intrigued, they obtained the brains of pigs processed for food production to study how widespread this postmortem viability might be in the intact brain. Four hours after the pig’s death, they connected the vasculature of the brain to circulate a uniquely formulated solution they developed to preserve brain tissue, utilizing a system they call BrainEx. They found neural cell integrity was preserved, and certain neuronal, glial, and vascular cell functionality was restored.
The new system solves the inability to apply certain techniques to study the structure and function of the intact large mammalian brain — which hinders rigorous investigations into topics like the roots of brain disorders, as well as neuronal connectivity in both healthy and abnormal conditions.
The brains of humans and other mammals are highly vulnerable to interruptions in blood flow and decreases in oxygen levels. Here we describe the restoration and maintenance of microcirculation and molecular and cellular functions of the intact pig brain under ex vivo normothermic conditions up to four hours post-mortem. We have developed an extracorporeal pulsatile-perfusion system and a haemoglobin-based, acellular, non-coagulative, echogenic, and cytoprotective perfusate that promotes recovery from anoxia, reduces reperfusion injury, prevents oedema, and metabolically supports the energy requirements of the brain. With this system, we observed preservation of cytoarchitecture; attenuation of cell death; and restoration of vascular dilatory and glial inflammatory responses, spontaneous synaptic activity, and active cerebral metabolism in the absence of global electrocorticographic activity. These findings demonstrate that under appropriate conditions the isolated, intact large mammalian brain possesses an underappreciated capacity for restoration of microcirculation and molecular and cellular activity after a prolonged post-mortem interval.