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        Biochemical and functional changes of rat liver spheroids during spheroid formation and maintenance in culture. ii. nitric oxide synthesis and related changes

        Author
        Xu, J.
        Ma, M.
        Purcell, W.
        Attention
        2299/2481
        Abstract
        Liver cells isolated from intact tissue can reaggregate to form three-dimensional, multicellular spheroids in vitro. During this process, cells undergo a histological and environmental change. How cells respond biochemically to this change has not been studied in detail previously. We have investigated some biochemical changes in rat liver cells during the formation and maintenance of spheroids. Liver cells were isolated from male Sprague rats and spheroids cultured by a gyrotatory-mediated method. Liver cells were shown to respond to the isolation procedure and the formation of spheroids triggered histological environmental changes that increased arginine uptake, nitric oxide (NO) and urea syntheses, as well as raised levels of GSH, GSSG, glutamic acid and aspartic acid secretion within the first couple of days after cell isolation. Levels were maintained at a relatively stable level in the mature spheroids (>5 days) over the 3 week period of observation. P450 1A1 activity was lost in the first 2 days and gradually recovered thereafter. This study, for the first time, shows that liver cells after isolation and during spheroid formation actively uptake arginine and increase NO and urea syntheses. A high level of NO is likely to play an important role in modulating a series of biochemical changes in liver cells. It is considered that liver cells actively respond to the ‘challenge’ induced by the isolation procedure and subsequent histological environmental changes, and biochemical modulation and instability result. The stable cell–cell contacts and histological environment in mature spheroids permit and support functional recovery and maintenance in vitro. This period of stability permits the use of spheroids in toxicity studies to establish acute and chronic paradigms. © 2003 Wiley-Liss, Inc.
        Publication date
        2003
        Published in
        Journal of Cellular Biochemistry
        Published version
        https://doi.org/10.1002/jcb.10731
        Other links
        http://hdl.handle.net/2299/2481
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