Cell proliferationFeeder cellsMitomycin CFeeder cells play important roles in In-vitro culture of stem cells. However, the preparation protocol of feeder cells produced by bovine embryonic fibroblast cells (bEFs) is still lack. In this study, the preparation of bEF-feeder by Mitomycin C was ...
Non-small cell lung cancer (NSCLC) cells were pre-treated with mitomycin C (MMC) and then co-cultured with peripheral blood mononuclear cells (PBMCs) to investigate the effect of the combination of MMC with PD-L1 antibody. The drug combination was also evaluated in vivo in Lewis lung ...
11.)Trypsinize the epithelial cells using their normal protocol (for keratinocytes trypsin 2 minutes RT, aspirate off the trysin and then incubate for 5 min at 37oC, tap plate, resuspend in DMEM/F12). Epithelial cells should be plated back onto mitomycin C treated feeder layers in low densi...
The combination of mitomycin C and rapamycin inactivated S6K1, not Akt, and resulted in depho- sphorylation of Bad, which inactivated its pro-apoptotic function, inducing Bak oligomerization and mitochondrial- dependent apoptotic cell death. Results The combination of mitomycin C and rapamycin ...
2.6. Cell cycle analysis The cell cycle distribution was examined using a FITC BrdU Flow Kit (BD Pharmingen, USA), according to the manufacturer’s protocol. A flow cytometer (Flow Sight, Merck, Darmstadt, Germany) was used to assess DNA content and the cell cycle distribution. 2.7. Analysis...
Cell death was measured by acridine orange/ethidium bromide (AO/EB) staining, subG1 analysis using PI/RNAse staining buffer (cat-no. 550825, BD Pharmingen™) and annexin-V/7-AAD staining (cat-no. 556547, BD Pharmingen™) according to manufacturer's protocol. Mitochondrial membrane potential...
HeLa, SiHa and THP-1 MΦ were plated in 35 mm culture dishes. After 24 h, cells were pretreated with GW9662 (10 μM) for 2 h. Thereafter, MMC (500 nM) treatment was given and cells were incubated for 22 h. Whole-cell lysates were subjected to western blottin...
Using 4 human cancer cell lines, 4 tumors xenografted into nude mice, and 11 fresh tumor specimens removed at surgery, we investigated the relevance of NAD(P)H:quinone oxidoreductase (DT-diaphorase, DTD) activity (nmoles/min/mg protein) to mitomycin C (MMC)-induced cytotoxicity. In culture ...
HeLa, SiHa and THP-1 MΦ were plated in 35 mm culture dishes. After 24 h, cells were pretreated with GW9662 (10 μM) for 2 h. Thereafter, MMC (500 nM) treatment was given and cells were incubated for 22 h. Whole-cell lysates were subjected to western blotting for FasL. (c) ...
The level of DNA damage in endothelial cells was assessed using the cytokinesis-block micronucleus assay (CBMN assay) according to the standard protocol [80] with modifications described previously [17]. Cell growth media was removed from each well of 6-well plates; cells were washed with ice-...