Each passage, cells were seeded at similar cell numbers (20% confluency) and carried until 80% confluent at which point they were passaged again. After 100 generations (24 passages), each cell line was single-cell cloned and two of each clone (WT pool, WT Clone 3, HMCES-/- Clones ...
Influence of senescent cells on the moving trajectories of normal cells in confluency. (a) A snapshot image of a CPM simulation: (non-active) senescent cells (red) and (active) normal cells (yellow). One normal cell (randomly selected; marked by block dot) is traced in the right-hand s...
Each passage, cells were seeded at similar cell numbers (20% confluency) and carried until 80% confluent at which point they were passaged again. After 100 generations (24 passages), each cell line was single-cell cloned and two of each clone (WT pool, WT Clone 3, HMCES-/- Clones ...
Place your sterile and coated coverslips into a new sterile 24-well culture plate. Split your adherent cell line as you normally would with growth media. Plate your cells at your normal confluency (~10%) onto the surface of your coverslips. It will take ~400 to 500 µl of media to co...
Result Comparative analysis of VOCs in the human lung cancer and normal lung cell lines To extract the VOCs, upon reaching confluency the cell lines were grown for one, two and three weeks. After a one week incubation period, no dead cells were observed in any of the cell lines. After ...
MSCs from passage 5–8 were seeded at 4000 cells/cm2 and cultured for 7 days in MSC media up to 50–70% confluency, washed twice with PBS, and the media changed to OptiMEM with no serum (Invitrogen, Massachusetts, USA). After two days, the supernatant was collected, centrifuged at 700...
Upon reaching confluency, the cells were passaged onto a 10 cm dish precoated with Geltrex (Gibco, ThermoFisher Scientific). The following day, culture media was changed to StemFlex media (StemFlex Medium, Gibco, ThermoFisher Scientific). Media was then changed every 1–2 days until stable ...
Cells were subcultured when confluency reached 80–90%. Cells between 3rd to 5th passages were used in the following experiments. RNA synthesis The linear dsDNA of firefly luciferase (FLuc), GFP, VACC E3, mHGF and mCXCL12 were obtained from Integrated DNA Technologies (Supplementary Table 1)....
In use of the invention, an initial sort may be performed on the basis only of the first marker, e.g. SDC2 expression. This method generally also isolates unwanted cells, meaning cells that are not stromal stem cells and may be e.g. B-cells or T-cells. By the further step of isolat...
Cells were then replated at 1×105 cells/mL. Cultures that were allowed to achieve confluency where found to have diminished capacity for both proliferation and differentiation. Subsequent to this finding, cultures were not allowed to achieve higher densities than 1×106 cells/75 cm2. Example ...