Department of Physiology and Immunology, Faculty of Biology, University of Barcelona, Av. Diagonal 643, Barcelona 08028, Spain.
Domest Anim Endocrinol. 2013 Jul;45(1):1-10. doi: 10.1016/j.domaniend.2013.02.002. Epub 2013 Mar 16.
A preadipocyte primary cell culture was established to gain knowledge about adipose tissue development in gilthead sea bream (Sparus aurata), one of the most extensively produced marine aquaculture species in the Mediterranean. The preadipocytes obtained from the stromal-vascular cell fraction of adipose tissue proliferated in culture, reaching confluence around day 8. At that time, the addition of an adipogenic medium promoted differentiation of the cells into mature adipocytes, which showed an enlarged cytoplasm filled with lipid droplets. First, cell proliferation and differentiation were analyzed under control and adipogenic conditions during culture development. Next, the effects of insulin, GH, and IGF-I on cell proliferation were evaluated at day 8. All peptides significantly stimulated proliferation of the cells after 48 h of incubation (P < 0.002 for GH and IGF-I and P < 0.05 for insulin), despite no differences were observed between the different doses tested. Subsequently, the effects of insulin and IGF-I maintaining differentiation when added to growth medium were studied at day 11, after 3 d of induction with adipogenic medium. The results showed that IGF-I is more potent than insulin enhancing differentiation (P < 0.01 for IGF-I compared with the control). In summary, a primary culture of gilthead sea bream preadipocytes has been characterized and the effects of several regulators of growth and development have been evaluated. This cellular system can be a good model to study the process of adipogenesis in fish, which may help improve the quality of the product in aquaculture.
建立了原代前体脂肪细胞培养体系,以了解地中海地区最广泛养殖的海水养殖鱼类之一——真鲷(Sparus aurata)的脂肪组织发育。从前体脂肪细胞中获得的基质血管细胞分数在培养中增殖,大约在第 8 天达到汇合。此时,添加脂肪生成培养基可促进细胞分化为成熟的脂肪细胞,这些细胞的细胞质增大,充满了脂滴。首先,在培养过程中分析了对照和脂肪生成条件下的细胞增殖和分化。然后,在第 8 天评估了胰岛素、GH 和 IGF-I 对细胞增殖的影响。所有肽在孵育 48 小时后都显著刺激了细胞的增殖(GH 和 IGF-I 的 P<0.002,胰岛素的 P<0.05),尽管在测试的不同剂量之间没有观察到差异。随后,在添加到生长培养基中时,研究了胰岛素和 IGF-I 在维持分化方面的作用,这是在诱导脂肪生成培养基 3 天后的第 11 天进行的。结果表明,IGF-I 比胰岛素更能增强分化(与对照相比,IGF-I 的 P<0.01)。总之,已对真鲷前体脂肪细胞的原代培养进行了表征,并评估了几种生长和发育调节剂的作用。该细胞体系可以作为研究鱼类脂肪生成过程的良好模型,这可能有助于提高水产养殖产品的质量。