| Title |
Towards a more representative in vitro method for fish ecotoxicology: morphological and biochemical characterisation of three-dimensional spheroidal hepatocytes
|
|---|---|
| Published in |
Ecotoxicology, June 2012
|
| DOI | 10.1007/s10646-012-0965-5 |
| Pubmed ID | |
| Authors |
Matthew G. Baron, Wendy M. Purcell, Simon K. Jackson, Stewart F. Owen, Awadhesh N. Jha |
| Abstract |
The use of fish primary cells and cell lines offer an in vitro alternative for assessment of chemical toxicity and the evaluation of environmental samples in ecotoxicology. However, their uses are not without limitations such as short culture periods and loss of functionality, particularly with primary tissue. While three-dimensional (spheroid) technology is now established for in vitro mammalian toxicity studies, to date it has not been considered for environmental applications in a model aquatic species. In this study we report development of a reproducible six-well plate, gyratory-mediated method for rainbow trout (Oncorhynchus mykiss) hepatocyte spheroid culture and compare their functional and biochemical status with two-dimensional (2D) monolayer hepatocytes. Primary liver spheroid formation was divided into two stages, immature (1-5 days) and mature (≥6 days) according to size, shape and changes in functional and biochemical parameters (protein, glucose, albumin and lactate dehydrogenase). Mature spheroids retained the morphological characteristics (smooth outer surface, tight cell-cell contacts) previously described for mammalian spheroids as demonstrated by light and scanning electron microscopy. Glucose production and albumin synthesis were significantly higher in mature spheroids when compared to conventional 2D monolayer cultures (P < 0.01) and increased as spheroids matured (P < 0.01). Basal lactate dehydrogenase (LDH) leakage significantly decreased during spheroid formation and was significantly lower than 2D cultures (P < 0.01). It is therefore suggested that mature spheroids can maintain a high degree of functional, biochemical and morphological status over-time in culture that is superior to conventional 2D models and can provide realistic organotypic responses in vitro. Trout spheroids that take ~6-8 days to reach maturity would be suitable for use in acute toxicological tests and since it is possible to culture individual spheroids for over a month, there is potential for this work to lead towards in vitro bioaccumulation alternatives and to conduct high throughput screens of chronic exposure. This is an important step forward for developing alternative in vitro tools in future fish ecotoxicological studies. |
Mendeley readers
The data shown below were compiled from readership statistics for 87 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Malaysia | 1 | 1% |
| Brazil | 1 | 1% |
| Canada | 1 | 1% |
| Denmark | 1 | 1% |
| Spain | 1 | 1% |
| Unknown | 82 | 94% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 19 | 22% |
| Student > Master | 17 | 20% |
| Student > Ph. D. Student | 16 | 18% |
| Student > Bachelor | 5 | 6% |
| Professor | 4 | 5% |
| Other | 13 | 15% |
| Unknown | 13 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 30 | 34% |
| Environmental Science | 14 | 16% |
| Biochemistry, Genetics and Molecular Biology | 6 | 7% |
| Chemistry | 4 | 5% |
| Pharmacology, Toxicology and Pharmaceutical Science | 3 | 3% |
| Other | 10 | 11% |
| Unknown | 20 | 23% |
Attention Score in Context
This research output has an Altmetric Attention Score of 4. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 18 December 2014.
All research outputs
#6,915,761
of 22,681,577 outputs
Outputs from Ecotoxicology
#217
of 1,469 outputs
Outputs of similar age
#49,377
of 164,445 outputs
Outputs of similar age from Ecotoxicology
#4
of 14 outputs
Altmetric has tracked 22,681,577 research outputs across all sources so far. This one has received more attention than most of these and is in the 68th percentile.
So far Altmetric has tracked 1,469 research outputs from this source. They receive a mean Attention Score of 3.5. This one has done well, scoring higher than 84% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 164,445 tracked outputs that were published within six weeks on either side of this one in any source. This one has gotten more attention than average, scoring higher than 68% of its contemporaries.
We're also able to compare this research output to 14 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 71% of its contemporaries.