Primary hepatocytes as an useful bioassay to characterize metabolism and bioactivity of illicit steroids in cattle

Isolation and cultivation of adult primary bovine hepatocytes from abattoir derived liver

“…This result was slightly lower compared to Giantin et al (2012[3]) who achieved 88 % viable cells and who also obtained liver cells from two heifers after slaughter or to Panda et al (2015[9]) who obtained 82 % viable cells from buffalo liver after slaughter. It is known from rat hepatocytes that after 72 h of culture on collagen monolayer, the cells develop perturbed cell morphology, and cells spread out and form fibroblast-like protrusions.…”

“…These results have also been confirmed after 48 h in monolayer cultures of bovine hepatocytes (Tuschl et al, 2009[14]; Tuschl and Mueller, 2006[15]) and in the present study. In contrast to Giantin et al (2012[3]), in the present study, bovine hepatocytes were cultured after isolation in a sandwich culture system. The polygonal cell shape of the cells was still stable after 14 days, and this result is comparable to results obtained from rat hepatocytes cultured between two layers of collagen (Tuschl and Mueller, 2006[15]).…”

“…Donkin and Armentano (1993[1]) have described a process for the culturing of primary bovine hepatocytes up to 41 h after separation from male calves. Giantin et al (2012[3]) described a method for isolating primary hepatocytes from the caudate lobe after slaughter in cattle. They have conducted viable cell-cultures from these cells for 24 h in a monolayer system.…”

In Vitro Androgen Bioassays as a Detection Method for Designer Androgens

“…These androgens or androgen-like molecules enhance the growth of the animals and are used as a means to increase profit. Naturally occurring steroids such as testosterone or synthetic androgens such as 19-nortestosterone, trenbolone acetate, and medroxyprogesterone are used to illicitly augment growth of animal livestock [42,43]. In addition, prohormones such as dehydroepiandrosterone (DHEA) are being used, and these are often hard to detect if they have been exogenously administered [44].…”

“…GC-MS may also fail to detect novel structures or those with unknown metabolic profiles. Bioassays can complement the screening of samples as they are capable of detecting hormonally active compounds in prepared extracts and if the appropriate host cell is used for the bioassay such as hepatocytes then such assays may also detect prohormones [43,45,46]. For these reasons, bioassays are ideal for the detection of androgens or proandrogens added to nutritional supplements.…”