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Cells were cultured at sub-confluence and utilized between passages 5 and 10.
Fresh MSCs at passages 5 to 7 were used for the present study.
Vascular smooth muscle cells (between passages 5 and 10) were incubated with 0.1% neonatal calf serum for 48 h, and then treated with angiotensin II (100 nM) in the absence or presence of CGRP for 24 h.
Early passages (5 to 9) of cells grown to 80-90% confluence were used for all experiments.
Following adaptation in quail through serial lung passages (5 times), the adapted virus gained transmission properties in quail and replication/transmission properties in chickens [24].
Experiments were performed using cells at passages 5 to 9. All fibroblasts (WT n = 3; NBCCS n = 6) were cultured in a tridimensional type I collagen matrix called dermal equivalent.
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Passages 5 15 were used in this study.
Total RNA was isolated from cell cultures of the clones (passages 5 7) using RNAeasy midi kits (Qiagen sciences, Maryland).
Explant technique was used to isolate primary fibroblasts as described previously [33] and fibroblasts were used between passages 5 9 and maintained as described above.
In this work we characterised C5 early (passages 5 7) (C5E), C5 late (passages 20 30) (C5L), and B10 late (passages 30 60) (B10L) cell lines.
Experiments were done on C5 early (passages 5 7) (C5E), C5 late (passages 20 30) (C5L), and B10 late (passages 30 60) (B10L) cell lines.
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