Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/104686
Title: Quantitative analysis of rat adipose tissue cell recovery, and non-fat cell volume, in primary cell cultures.
Author: Rotondo, Floriana
Romero, María del Mar
Ho-Palma, Ana Cecilia
Remesar Betlloch, Xavier
Fernández López, José Antonio
Alemany, Marià, 1946-
Keywords: Teixit adipós
Cèl·lules
Rates (Animals de laboratori)
Adipose tissues
Cells
Rats as laboratory animals
Issue Date: 29-Nov-2016
Publisher: PeerJ
Abstract: Background. White adipose tissue (WAT) is a complex, diffuse, multifunctional organ which contains adipocytes, and a large proportion of fat, but also other cell types, active in defense, regeneration and signalling functions. Studies with adipocytes often require their isolation from WAT by breaking up the matrix of collagen fibres; however, it is unclear to what extent adipocyte number in primary cultures correlates with their number in intact WAT, since recovery and viability are often unknown. Experimental Design. Epididymal WAT of four young adult rats was used to isolate adipocytes with collagenase. Careful recording of lipid content of tissue, and all fraction volumes and weights, allowed us to trace the amount of initial WAT fat remaining in the cell preparation. Functionality was estimated by incubation with glucose and measurement of glucose uptake and lactate, glycerol and NEFA excretion rates up to 48 h. Non-adipocyte cells were also recovered and their sizes (and those of adipocytes) were measured. The presence of non-nucleated cells (erythrocytes) was also estimated. Results. Cell numbers and sizes were correlated from all fractions to intact WAT. Tracing the lipid content, the recovery of adipocytes in the final, metabolically active, preparation was in the range of 70 75%. Cells showed even higher metabolic activity in the second than in the first day of incubation. Adipocytes were 7%, erythrocytes 66% and other stromal (nucleated cells) 27% of total WAT cells. However, their overall volumes were 90%, 0.05%, and 0.2% of WAT. Non-fat volume of adipocytes was 1.3% of WAT. Conclusions. The methodology presented here allows for a direct quantitative reference to the original tissue of studies using isolated cells. We have also found that the ``live cell mass'' of adipose tissue is very small: about 13 mL/g for adipocytes and 2 mL/g stromal, plus about 1 mL/g blood (the rats were killed by exsanguination). These data translate (with respect to the actual ``live cytoplasm'' size) into an extremely high metabolic activity, which make WAT an even more significant agent in the control of energy metabolism.
Note: Reproducció del document publicat a: https://doi.org/10.7717/peerj.2725
It is part of: PeerJ, 2016, vol. 4, p. e2725
Related resource: https://doi.org/10.7717/peerj.2725
URI: http://hdl.handle.net/2445/104686
ISSN: 2167-8359
Appears in Collections:Articles publicats en revistes (Bioquímica i Biomedicina Molecular)

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