1 2 3 4

		Two methods of limb bud culture exist. Chorioallantioc membrane (CAM) grafting is a culture method commonly used in the past. In this type of experiment, the severed limb bud is grafted into a host egg on a Y- shaped junction of blood vessels. The limb grows on nutrients supplied by the host egg. A more recent application of limb bud uses no host but a growth medium in vitro. The culture medium contains elements essential for continued growth and antibiotics to protect against infection. Although CAM grafts have been used in the past, an obvious advantage of an in vitro method is that manipulative agents, such as growth factors and hormones, can be added to the culture medium. The primary purpose of this experiment is to observe limb development by studying the presence of cartilage in cultured limbs. The secondary purpose is to investigate whether or not in vitroculture can support limb development. The control will be limbs that develop in ovo. Major Observations Limbs developed in vitroand cartilage was observed using Alcian green staining. In some cases, development had progressed to the point at which cartilage began changing into bone, and, thus, was not seen in the staining. Compared to control limbs, limbs grown in media were not as large or organized as well.