| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Articles |
a Department of Veterinary Anatomy and Public Health, Texas A&M University, College Station, Texas 77843
b Department of Cell Biology and Neuroscience and Department of Pathology,
c University of Texas Southwestern Medical School, Dallas, Texas 75235
d Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu, Hawaii 96822
For many infertile couples, intracytoplasmic germ cell/spermatozoon injection into unfertilized eggs may be their only hope for producing their own biological children. Thus far, success with injection of pre-spermatozoan germ cells such as round spermatids has not been as great as that of spermatozoon injection. This could be due in part to the difficulty of identifying younger (less mature) male germ cells in testicular biopsy dispersions. To improve the identification of various types of live, dispersed, human testicular cells in vitro, a comparative study of the morphological characteristics of human spermatogenic germ cells in vitro or in situ within seminiferous tubules was conducted. Live human testicular tissue was obtained from an organ-donating, brain-dead person with a high density of various germ cells. A cell suspension was obtained by enzymatic digestion, and cells were cultured for 3 days in an excessive volume (100-fold medium:cells; v:v) of HEPES-TC 199 medium at 5°C and observed live with Nomarski optics (interference-contrast microscopy). For comparative purposes, testes from ten men obtained at autopsy were fixed, embedded in epoxy resin, sectioned at 20 µm, and observed unstained by Nomarski optics. This approach allowed comparison of morphological characteristics of individual germ cells seen in vitro or in situ in the human testis. In both live and fixed preparations from control men with varied daily sperm production rates, Sertoli cells have oval to pear-shaped nuclei with indented nuclear envelopes and large nucleoli, which makes their appearance distinctly different from germ cells. The size, shape, and chromatin pattern of nuclei, and the presence of meiotic metaphase figures, acrosomic vesicles/structures, tails, and/or mitochondria in the middle piece of germ cells are characteristically seen in live cells in vitro and in those cells observed in the fixed seminiferous tubules. 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 and technical staff in infertility clinics when selecting specific germ cells from a testicular aspirate or enzymatically digested biopsy.
2 Correspondence. FAX: 409 847 8981; ljohnson1{at}tamu.edu
This article has been cited by other articles:
|
|
 |
|
  V. Roulet, H. Denis, C. Staub, A. Le Tortorec, B. Delaleu, A.P. Satie, J.J. Patard, B. Jegou, and N. Dejucq-Rainsford Human testis in organotypic culture: application for basic or clinical research Hum. Reprod., June 1, 2006; 21(6): 1564 - 1575. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Ehmcke, J. Wistuba, and S. Schlatt Spermatogonial stem cells: questions, models and perspectives Hum. Reprod. Update, May 1, 2006; 12(3): 275 - 282. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. P. Mishra and C. Shaha Estrogen-induced Spermatogenic Cell Apoptosis Occurs via the Mitochondrial Pathway: ROLE OF SUPEROXIDE AND NITRIC OXIDE J. Biol. Chem., February 18, 2005; 280(7): 6181 - 6196. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Marcon and G. Boissonneault Transient DNA Strand Breaks During Mouse and Human Spermiogenesis:New Insights in Stage Specificity and Link to Chromatin Remodeling Biol Reprod, April 1, 2004; 70(4): 910 - 918. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Jagannathan, E. L. Punt, Y. Gu, C. Arnoult, D. Sakkas, C. L. R. Barratt, and S. J. Publicover Identification and Localization of T-type Voltage-operated Calcium Channel Subunits in Human Male Germ Cells. EXPRESSION OF MULTIPLE ISOFORMS J. Biol. Chem., March 1, 2002; 277(10): 8449 - 8456. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. V. Abraham-Peskir, E. Chantler, E. Uggerhoj, and J. Fedder Response of midpiece vesicles on human sperm to osmotic stress Hum. Reprod., February 1, 2002; 17(2): 375 - 382. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Johnson, C. Staub, W. B. Neaves, and R. Yanagimachi Live human germ cells in the context of their spermatogenic stages Hum. Reprod., August 1, 2001; 16(8): 1575 - 1582. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
