|Back Topic Categories Search Previous Abstract Next Abstract|
Larry Johnson1 *, William B. Neaves2 *, Jeffrey J. Barnard2 , Genevieve E. Keillor1 , Steven W. Brown1 , Ryuzo Yanigimachi3 *
Texas A&M University, College Station, Texas 77843 1
University of Texas Southwestern Medical School, Dallas, Texas 75235 2
University of Hawaii School of Medicine, Honolulu, Hawaii 96822 3
To enhance the identification of various types of live, dispersed, human testicular cells in vitro, a comparative study of the morphologic characteristics of human spermatogenic germ cells in vitro or in situ within seminiferous tubules of defined spermatogenic stages was conducted. Live human testicular tissue was obtained from an organ-donating, brain-dead person. A cell suspension was obtained by enzymatic digestion, and cultured cells were observed live with Nomarski optics. Testes from ten men were obtained at autopsy within ten hours of death, fixed in glutaraldehyde, further fixed in osmium, embedded in Epon, sectioned at 20 micrometers, and observed unstained by Nomarski optics. This approach allowed comparison of morphologic characteristics of individual germ cells seen in vitro or in situ in the human testis in the context of the spermatogenic stage. In both live and fixed preparations, Sertoli cells have oval to pear-shaped nuclei with indented nuclear envelopes and large nucleoli, which makes their appearance distinctly different from germ cells. For germ cells, size, shape, and chromatic pattern of nuclei, the presence of meiotic metaphase figures, acrosomic vesicles/structures, tails, and/or mitochondria in the middle piece are characteristically seen in live cells in vitro and those in the fixed seminiferous tubules, and they lead to identification of live germ cells in man. Hence, this comparative approach allows verification of the identity of individual germ cells seen in vitro and provides a checklist of distinguishing characteristics of live human germ cells to be used by scientists interested studying live cells of known steps in spermatogenic development.
This abstract is being presented on Monday, August 2 at 8:00 AM to 10:15 AM at CUB 2nd Floor Ballroom.