Native cotton fiber and in vitro cotton fiber that was induced from cotton ovule callus by suspension culture were observed using transmission electron microscope and scanning electron microscope. The ovule surface on the first day preanthesis was quite smooth. On the anthesis, it had a lot of protuberances. Two kinds of callus, smooth and rough were found. The microfibrils of callus was vertical to the cell long axis and they changed their orientations with the development of the in vitro cotton fiber: from the vertical to shallow spiral and then to parallel to the cell long axis. So was the native cotton fiber. It suggests that in vitro cotton fiber and native cotton fiber have similar development process. Compared with the ovule surface cell, most callus cells had smaller nuclear. During the development of the fiber, the plasm of native cotton fiber was denser than that of in vitro fiber, and it has more cellular organ than in vitro fiber. The cell wall of native cotton fiber was thicker and denser than that of the in vitro cotton fiber too. It suggests that the physiological activity of in vitro cotton fiber was less active than native cotton fiber.