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Materials and Methods:

Since the sixth day marks the formation of lung buds (Romanoff 1960), we isolated the lungs of 6-day-old chick embryos

by dissection in Howards Ringer solution. The isolated lungs were then placed on Transwell filters, which were immersed in 2 mL

of BGJb culture solution. We performed two different experiments. In the first experiment, we observed the affects of an FGF

inhibitor on isolated BGJb cultured chick lungs over a 3-day period. The control for this experiment was cultured in just BGJb

medium. This experiment was duplicated, but the second trial it was allowed to culture for 5 days. In the second experiment we

used isolated 6-day-old chick lungs in a BGJb culture and placed FGF-loaded heparin beads on the side of the lungs that do not

usually form buds (Weaver et al. 2000). The control for this experiment had heparin beads that were not loaded with anything

placed on the side of chick lungs that do not form buds. This was done to rule out whether the heparin by itself has the potential

to induce bud formation.



We observed after 3 and 5 days that new lung buds did not form in the FGF inhibited cultures. However in both the

control and in the FGF inhibited cultures, the lung buds that had already formed proceeded to further develop as normal.

Although, in the control culture the lungs grew the additional lung buds that they should normally develop (Figure 1).

In the second experiment, we observed new lung buds formation that were induced by the FGF-loaded heparin beads on

the side of the lungs that do not normally form buds. In the control, we observed normal lung formation, but there was no

induction by the plain heparin beads (Figure 2).

© Cebra-Thomas, 2001

Last Modified: 31 May 2001

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