Feather Bud Development

Introduction

Procedure

Results/Data

Discussion

 

Acknowledgements

Works Cited

Downloads

Additional Pictures

Procedure:


1. Obtain a six-well plate. In each well, pipette 1.8 ml of Hank’s Balanced Salt Solution 1X. Rest Transwell 3452 culture inserts (24mm diameter) over the wells containing the salt solution.


2. Wash eight eight-day-old chick eggs with 70% ethanol. Open the chick embryos as demonstrated in class (at the blunt end of the shell, where the air sac is located), using forceps. Record the stages of the embryos, making sure to keep tract of which embryo is at which stage as they are removed from their shells.


3. Empty the embryos into separate petri dishes and remove them from their yolks. Then place the isolated chick embryos, using a plastic spoon, into smaller petri dishes (60mm) containing Howard Ringers Solution.


4. Cut away the anterior portion (i.e. remove the head) of the embryo using either scissors or forceps. Make a shallow incision along the trunk of the embryo and peel off as large a por-tion of skin as possible. Place this skin segment onto an insert covering one of the wells filled with Hank’s solution. Make sure the ≤inside "portion of the skin is touching the disk (see DB Lab CD; Cebra-Thomas, 2001).


5. Place well plates with skin cultures into an incubator, set at 5% CO2 and 37æC. Observe development of skin cultures periodically over a weeks time, noting the appearance and development of feather buds. Record observations and photograph skin cultures at every observation point to determine the extent to which the skin develops.


6. Limbs from the embryos may also be placed in analogous culture conditions (same media) with the caveat of using non-collagen culture inserts so that the limbs won’t stick too closely to the insert. These too may be observed periodically to determine if there would be any further development of the limbs, or feather buds on the limbs.

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