RESUMO
Slow cooling leads to a passive dehydration of cells, whereas rehydration during warming reflects the active regain of functionality. The ability to modulate such an energy demanding process could be instrumental in optimizing the cryo-arrest of living systems. In the present study, various levels of hypertonic stress were used to disturb the water content of cells and to define the energy profiles of aquaporins and (Na(+) conducting) cation channels during rehydration. Na(+) import was found to be the rate-limiting step in water restoration, whereas aquaporins merely played a permissive role. Indeed, regulated Na(+) import was increased 2-fold following cryo-arrests, thus facilitating the osmotic rehydration of cells. Freezing temperatures increased cell viscosity with a remarkable hysteresis and viscosity was a trigger of cation channels. The peptide hormone vasopressin was a further activator of channels, increasing the viability of post-cryo cells considerably. Hence, the hormone opens the path for a novel class of cryo-protectants with an intrinsic biological activity.
Assuntos
Adaptação Fisiológica , Pontos de Checagem do Ciclo Celular , Resposta ao Choque Frio , Congelamento , Pressão Osmótica , Aquaporinas/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células HeLa , Células Hep G2 , Humanos , Vasopressinas/farmacologia , Viscosidade , Canais de Sódio Disparados por Voltagem/metabolismoRESUMO
We present a novel noninvasive technology for quality control in biobanking. We implemented a contactless optical in situ method with a remote detection unit. The method detects physical and chemical changes by emission spectroscopy. In the present study, ice formation in a vitrified sample is revealed by Raman scattering. The technology allows us to monitor sample quality during cold storage and to assess the sample state after preservation, storage, or transport without the need for thawing.
