Diffusion Chamber Studies of Rat Peripheral Leucocytes in the Presence of Phytohaemagglutinin

Abstract

SUMMARY Rat peripheral leucocytes have been cultured in diffusion chambers in an effort to achieve greater consistency in the response of lymphocytes to phytohaemagglutinin. The diffusion chamber was found to provide an in vivo environment which obviates drastic changes in pH and eliminates other factors that act as major deterrents to successful and optimal responses of lymphocytes to this mitogen in vifro. The system shows reliability and consistency in maintaining excellent cytologic and morphologic integrity of the cells in culture.

Autoradiographic Studies of Human Lymphocytes Cultured in Vivo

“…This system has yielded consistently good results when applied to rat lymphocytes which are difficult to maintain in vitro (2)(3)(4)(5). The relationship of this in vitro proliferation to in vivo function, remains to be elucidated.…”

“…The technique for culturing blood lymphocytes in diffusion chambers was previously described (2). The technique for culturing blood lymphocytes in diffusion chambers was previously described (2).…”

Growth Characteristics of the Shay Chloroleukemia in Diffusion Chambers

“…More recently, diffusion chambers have been employed to investigate the response of rodent and human leukocytes to phytoheniagglutinin (PHA) and to study the kinetics of PHA-stimulated lymphocyte proliferation using tritiated thymidine autoradiography (7)(8)(9). The latter studies involved the serial sacrifice of rats bearing intraperitoneal diffusion chambers harvested at specific time intervals after isotope administration.…”

“…Wistar rats and Swiss mice were anesthetized by Metafane inhalation ( Pittman-Moore, Indianapolis) and the chambers were introduced into the peritoneal cavity by laparotomy. Two milliliters of medium 199 were injected into the peritoneal cavity after closure of the wound to retard development of connective tissue lesions around the chambers (7,8).…”