In this work, the flask-based LPC generation was decoupled from the 96-Transwell-based final maturation into AT2-like cells. Although, hiPSC-derived lung epithelial cell models have been used for disease modelling45,48, they have so far never been miniaturized and applied to medium throughput ...
MPS culture To ensure the adipogenesis of loaded adipocyte progenitor cells, the initial stage of differentiation was pre-induced in a tissue culture flask. Differentiating iPSC-MSCs on day 4, which contain tiny lipid droplets in > 80% cells, were dissociated (TrypLE Express, Gibco) and cen...
Subculturing1. Remove and discard culture medium. 2. Add 2.0mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed. Note: Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. 3. Centrifuge cells at 1500...
Three days after culture, floating cells were harvested and transferred to a new T25 flask without coating. The cells were maintained in NS-A2 CTL medium (Nissui) supplemented with 1% PSG in the presence of 700 IU/mL IL2 (Miltenyi Biotec) for 2weeks to obtain activated NK cells for ...
Fresh myeloid differentiation media was added back to the flask at a volume equal to the volume harvested in order to replenish the T175 flask. Harvested cells were plated onto tissue-culture treated plastic 12-well plates at 1.0E6 cells per well and differentiated for 7 days or more to ...
For embryoid body (EB) formation, the iPSCs were dissociated into single cells using Accutase® and transferred into a standing 25cm2 flask for cell culture suspension in mTSeR1 supplemented with 10 μM Y-27632. After 24 h, the medium was changed to EB medium consisting of DMEM/F12, 20...
TGFβ1 is a growth factor commonly used for maintenance of primary microglia and of iPS-microglia-like cell cultures [42, 52], but not included in our culture. We stimulated pMac with recombinant human TGFβ1 for 24 h, to assess whether TGFβ1 could “rescue” expression of the ...
(Diii) MAP2-positive neurons (red) and GFAP-positive astrocytes (green) co-culture. (E) iPSC-derived 3D organoid culture in spinner flask. (Ei,Eii) Organoids at different time points of culturing. (Scales in 100 μm, bright-field and immunofluorescent staining images are taken from ...
It is critical that cell confluency be 60–80%, cell viability be at least 90%, and the growth rate be in mid-logarithmic phase prior to sub culturing. Note: The following procedures apply to adherent cultures in a T-75 cu...
iPSCs were digested when 60–70% confluence was reached using dispase and colony clumps were transferred to an ultra-low attachment cell culture flask (Corning) in hESC medium. On day 6, embryoid bodies (EBs) were placed on 0.1% gelatin coated coverslips in 24-wellplate and cultured with ...