Journal: Scientific Reports

Article Title: Preservation of epithelial progenitor cells from collagenase-digested oral mucosa during ex vivo cultivation

doi: 10.1038/srep36266

Figure Lengend Snippet: Comparison of original and modified protocols for the ex vivo cultivation of human oral mucosal epithelial cells (OMECs). Left panel: In the original protocol, oral epithelium was isolated from oral mucosa with dispase II. Suspended OMECs were then dissociated with trypsin-EDTA, re-suspended and cultivated on de-epithelialized amniotic membrane (AM) with 3T3 fibroblasts as a feeder layer, in SHEM containing fetal bovine serum (FBS). Right panel: In the modified protocol, oral mucosal tissues were treated with collagenase A. The resulting cell aggregates were grown on AM without a 3T3 feeder layer. In addition, FBS was replaced with PLTMax. Two days after seeding, the culture medium was changed to serum-free medium (EpiLife) to prevent the overgrowth of submucosal fibroblasts.

Article Snippet: Materials DMEM:F12 (1:1), Opti-MEM, EpiLife medium, , trypsin-EDTA, phosphate-buffered saline (PBS), gentamicin, amphotericin B, Lipofectamine 2000 and Alexa-Fluor-conjugated secondary IgG were purchased from Invitrogen (Carlsbad, CA).

Techniques: Modification, Ex Vivo, Isolation

Journal: BMC Cancer

Article Title: Role of intracellular and extracellular annexin A1 in migration and invasion of human pancreatic carcinoma cells

doi: 10.1186/1471-2407-14-961

Figure Lengend Snippet: Expression and localization of cleaved and secreted form of ANXA1 from MIA PaCa-2 and PANC-1 cells. Cellular compartments were obtained as described in Methods section. Total (T), membrane (M), EDTA Wash (EW), cytosolic (C), nuclear (N) and supernatant (S) ANXA1 expression in protein extracts from MIA PaCa-2 and PANC-1 was examined by Western blot with anti-ANXA1 antibody. The protein bands were normalized on tubulin levels. The data are representative of 5 experiments with similar results.

Article Snippet: Nuclear extracts MIA PaCa-2 and PANC-1 cells were washed twice with PBS, detached with trypsin-EDTA 1× in PBS (Euroclone), harvested in PBS and centrifuged for 5 minutes at 600 × g at 4°C.

Techniques: Expressing, Western Blot