title: Freezing behavior of adherent neuron-like cells and morphological change and viability of post-thaw cells.
abstract: Freezing of nerve cells forming a neuronal network has largely been neglected, despite the fact that the cryopreservation of nerve cells benefits the study of cells in the areas of medicine and poison screening. Freezing of nerve cells is also attractive for studying cell morphology because of the characteristic long, thread-like neurites extending from the cell body. In the present study, freezing of neuron-like cells adhering to the substrate (differentiated PC12 cells), in physiological saline, was investigated in order to understand the fundamental freezing and thawing characteristics of nerve cells with neurites. The microscopic freezing behavior of cells under different cooling rates was observed. Next, the post-thaw morphological changes in the cells, including the cytoskeleton, were investigated and post-thaw cell viability was evaluated by dye exclusion using propidium iodide. Two categories of morphological changes, beading and shortening of the neurites, were found and quantified. Also, the morphological changes of neurites due to osmotic stress from sodium chloride were studied to gain a better understanding of causation. The results showed that morphological changes and cell death were promoted with a decrease in end temperature during freezing.
authors: Department of Biological Functions and Engineering, Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, Wakamatsu, Kitakyushu, Fukuoka 808-0196, Japan.
authors: Makoto
authors: M
authors: Uemura
authors: Department of Biological Functions and Engineering, Graduate School of Life Science and Systems Engineering, Kyushu Institute of Technology, Wakamatsu, Kitakyushu, Fukuoka 808-0196, Japan. Electronic address: ishiguro@life.kyutech.ac.jp.
authors: Hiroshi
authors: H
authors: Ishiguro
mesh_terms: D000818:Animals; D016923:Cell Death; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003460:Crystallization; D003599:Cytoskeleton / Q000502:physiology; D005615:Freezing / Q000009:adverse effects*; D016501:Neurites / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D009997:Osmotic Pressure / Q000502:physiology*; D011419:Propidium; D051381:Rats; D012965:Sodium Chloride
chemical_list: D011419:Propidium; D012965:Sodium Chloride
keywords: Cell viability; Cytoskeleton; Freezing; Ice crystals; Microscopic behavior; Morphology; Nerve cells; Neurites; Thawing