Differentiation of salisphere into acinar cells. (A) Amylase expressing cells (AC) in submandibular gland tissue (Tissue) were also present at the onset of culture (A-D0), and were visualized in the sphere at the onset of day 5 (A-D5), whereas granulae-containing spheres appeared in culture at later time-points (A-D10). Antibody labeling is shown in brown, nuclei in blue. Scale bar = 50 mm. (D = duct cells, AC = acinar cells, D0–3– 5–10 represent days in culture). (B) Real time RT-PCR confirmed the enhanced expression of amylase during in vitro culturing and differentiation. Error bars represent SEM (N = 2). Amylase mRNA expression levels at 2 days of culture were normalized to one. Reproduced with permission from: .