Volumes used in this protocol are for 75 cm 2 flasks; proportionally reduceor increase amount of dissociation medium for culture vessels of other sizes. Flasks do not become 100% confluent. Cells are rounded and have a tendency to float in the medium. ...
regulationoftelomeraseactivityinhumanleukamiacellK562withinterferona.Methods:ThehumanleukamiacellK562inculturewastreatedbyinterferona-2b.Byusingtelomeraserepeatamplificationprotocol(TRAP)combinedwithELISA,thetelomeraseactivityinhumanleukamiacellK562beforeandafterthetreatmentwasdetected.Byusingdirectedcountingwithbloodcell...
Therefore, coculture systems of irradiated feeder cells and NK cells in media containing IL-2 and IL-15 have been developed to generate large numbers of NK cells, although NK cell expansion protocol using anti-CD3 antibody (OKT-3) without feeder cells has also been developed. Commonly used ...
Cell cultures subjected to serum deprivation and hydroxyurea (HU) treatment demonstrated less than 5% of the cells to be in S phase. Four hours after release from HU, 63% of the cells were in S phase, as detected by immunofluorescence. This protocol offers a method for synchronization of ...
Cell culture K562 cell line was cultured in RPMI 1640 medium (GIBCO, Life Technologies) supplemented with 10% (v/v) fetal bovine serum, 100 units/ml Penicillin, and 100 μg/ml Streptomycin (P/S) (GIBCO, Life Technologies) in incubator (5% CO2, at 37°C). ...
present in all cell types, and are rapidly released into the cell culture medium upon plasma membrane damage. The obtained OD value reflects the amount of residue of live cells with an intact plasma membrane, because the cells, instead of the liquid phase of cell culture, were used in the ...
culture medium (from 0.1 to 2µM) for 9 months, as previously described (23). The concentration of imatinib was increased by 0.1µM at each step and was maintained for 15-30 days, depending on the proportion of surviving cells. The resulting K562IRcell line was resistant to 2µM...
Cell apoptosis assay The Annexin V-FITC apoptosis detection kit I was used to detect and quantify apoptosis by flow cytometry according to the manufacturer’s protocol. In brief, treated and untreated K562 cells were prepared as follows: A total of 5 × 105 cells were collected, washed with...
A negative ‘no template MAPK pathway is instrumental in cell growth, differentiation and control’ was included for each primer pair. The dissociation mutations in its downstream effectors are related to numerous protocol was performed at the end of each run to check for non- cancers [12]. In...
In the case of K562/A02 cell culture, ADR (1 μg/ml, final concentration) was added in the medium to maintain the MDR phenotype. The cells were further cultured in ADR-free medium for 2 weeks before experiment.Cell proliferation assay Cell proliferation assay was performed using MTT assay ...