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Fig. 3 | BMC Biotechnology

Fig. 3

From: Continuous long-term cytotoxicity monitoring in 3D spheroids of beetle luciferase-expressing hepatocytes by nondestructive bioluminescence measurement

Fig. 3

3D culture of primary hepatocytes isolated from CAG-ELuc/MI-MAC Tc mice. Primary hepatocytes were seeded in the 96-well format Cell-ableTM, in which feeder cells (3T3-Swiss albino) were pre-seeded 1 day before hepatocyte seeding, at the density of 4 × 104 cells/well. The culture medium containing 300 μM D-luciferin was refreshed every 2 to 3 days. a Morphology (left panel) and glycogen accumulation (right panel) of a 3D spheroid on culture day 7. Glycogen was stained with PAS. Scale bar indicates 50 μm. b Sequential changes of albumin secretion from 3D spheroids. Albumin concentration in the culture medium on the indicated culture day was measured by ELISA, and divided by culture day. Error bars indicate standard deviations (n = 8). c Sequential changes of ELuc bioluminescence from 3D spheroids. Bioluminescence from the 3D spheroids shown in (b) was measured nondestructively for 3 sec in each well using a luminometer. Error bars indicate standard deviations (n = 8)

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BMC Biotechnology

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