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Prostaspheres were cultured in Millicell hanging cell culture inserts with 1.0 µm PET transparent membranes (Millipore) on 6-well plates (Costar).
Polyethylene terephthalate membrane (8 μm pore size) Millicell hanging cell culture inserts were purchased from Millipore.
For LX2 cell coculture, 6-well 0.4 μm Millicell hanging cell culture inserts (Switzerland) were used.
Generally, 5 × 10 Huh7 cells were placed in 6-well plate while 1 × 10 LX2 cells were placed in hanging cell inserts.
The cell invasion assay was performed using a polyethylene terephthalate membrane (8 μm pore size) with Millicell hanging cell culture inserts and BD Matrigel.
Three days after isolation, BCECs reached about 80% confluence and were passaged onto collagen type IV- and fibronectin-coated 12 well polyethylene terephthalate, 1.0 µm hanging cell culture inserts at a cell density of 1 × 10 cells/cm.
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In this study, the volume of each hanging drop (cell density: 1.5 × 10 cells mL−1) was 38 μL.
COS cells were harvested 24 hours later and aggregates were made by hanging 105 COS cells in a 20 µl drop for 24 hours.
For initiation of embryoid body formation in hanging drops cells were dissociated by 0.05% trypsin and 0.04% EDTA in PBS and collected by centrifugation.
On day 0, we aggregated ES cells (5 × 10 cells/mL) by culturing in 20-μL hanging drops in cell culture medium without LIF.
He was found hanging in his cell in Hassockfield Secure Training Centre (STC), County Durham, hours after he was restrained by staff.
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