| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
Journal of Andrology, Vol 17, Issue 6 708-717, Copyright © 1996 by The American Society of Andrology
JOURNAL ARTICLE |
A. R. Morena, C. Boitani, M. Pesce, M. De Felici and M. Stefanini
Institute of Histology and General Embryology, University of Rome La Sapienza, Italy.
We have developed a new method that allows isolation of highly purified type A spermatogonia from prepubertal rats. The procedure is based on the maximal release of spermatogonia from the seminiferous epithelium obtained by the complete enzymatic digestion of the tubular basal lamina, followed by removal of contaminating somatic cells through adhesion to plastic dishes coated with the lectin Datura stramonium agglutinin and fractionation on a discontinuous Percoll gradient. The cell suspension obtained contains up to 85% type A spermatogonia. Besides morphological criteria, the identification of germ cells and somatic cells has been performed by means of immunocytochemical markers, such as c-kit receptor, which is present only in germ cells, and vimentin, which is present only in somatic cells. All type A spermatogonia isolated were c-kit positive, thus suggesting that c-kit receptor is present in both undifferentiated and differentiating type A spermatogonia. Preliminary culture experiments demonstrate that spermatogonia survival in vitro was significantly improved by the addition of 10% fetal calf serum or horse serum to the culture medium; however, optimal culture conditions remain to be established. In vitro studies on isolated spermatogonia may provide a significant contribution toward elucidation of the mechanisms regulating spermatogonial proliferation and differentiation.
This article has been cited by other articles:
 |
|
 |
|
  M. Monticone, L. Tonachini, S. Tavella, P. Degan, R. Biticchi, F. Palombi, R. Puglisi, C. Boitani, R. Cancedda, and P. Castagnola Impaired expression of genes coding for reactive oxygen species scavenging enzymes in testes of Mtfr1/Chppr-deficient mice Reproduction, September 1, 2007; 134(3): 483 - 492. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Raucci and M. M. Di Fiore The c-kit receptor protein in the testis of green frog Rana esculenta: seasonal changes in relationship to testosterone titres and spermatogonial proliferation Reproduction, January 1, 2007; 133(1): 51 - 60. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Khaira, D. McLean, D. A. Ohl, and G. D. Smith Spermatogonial Stem Cell Isolation, Storage, and Transplantation J Androl, July 1, 2005; 26(4): 442 - 450. [Full Text] [PDF]
|
|
|
|
 |
|
 N. Sofikitis, E. Pappas, A. Kawatani, D. Baltogiannis, D. Loutradis, N. Kanakas, D. Giannakis, F. Dimitriadis, K. Tsoukanelis, I. Georgiou, et al. Efforts to create an artificial testis: culture systems of male germ cells under biochemical conditions resembling the seminiferous tubular biochemical environment Hum. Reprod. Update, May 1, 2005; 11(3): 229 - 259. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Jarvis, D. J. Elliott, D. Morgan, R. Winston, and C. Readhead Molecular markers for the assessment of postnatal male germ cell development in the mouse Hum. Reprod., January 1, 2005; 20(1): 108 - 116. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Chiarella, R. Puglisi, V. Sorrentino, C. Boitani, and M. Stefanini Ryanodine receptors are expressed and functionally active in mouse spermatogenic cells and their inhibition interferes with spermatogonial differentiation J. Cell Sci., August 15, 2004; 117(18): 4127 - 4134. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Izadyar, J. J. Matthijs-Rijsenbilt, K. D. Ouden, L. B. Creemers, H. Woelders, and D. G. de Rooij Development of a Cryopreservation Protocol for Type A Spermatogonia J Androl, July 1, 2002; 23(4): 537 - 545. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. M. van Pelt, H. L. Roepers-Gajadien, I. S. Gademan, L. B. Creemers, D. G. de Rooij, and F. M. F. van Dissel-Emiliani Establishment of Cell Lines with Rat Spermatogonial Stem Cell Characteristics Endocrinology, May 1, 2002; 143(5): 1845 - 1850. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Roelants, F. Schneider, F. Goritz, J. Streich, and S. Blottner Seasonal Changes of Spermatogonial Proliferation in Roe Deer, Demonstrated by Flow Cytometric Analysis of c-kit Receptor, in Relation to Follicle-Stimulating Hormone, Luteinizing Hormone, and Testosterone Biol Reprod, February 1, 2002; 66(2): 305 - 312. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Fragale, R. Puglisi, A. R. Morena, M. Stefanini, and C. Boitani Age-dependent activin receptor expression pinpoints activin A as a physiological regulator of rat Sertoli cell proliferation Mol. Hum. Reprod., December 1, 2001; 7(12): 1107 - 1114. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Goddard, S. Bauer, A. Gougeon, F. Lopez, N. Giannetti, C. Susini, M. Benahmed, and S. Krantic Somatostatin Inhibits Stem Cell Factor Messenger RNA Expression by Sertoli Cells and Stem Cell Factor-Induced DNA Synthesis in Isolated Seminiferous Tubules Biol Reprod, December 1, 2001; 65(6): 1732 - 1742. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. H.G.J. Schrans-Stassen, P. T.K. Saunders, H. J. Cooke, and D. G. de Rooij Nature of the Spermatogenic Arrest in Dazl -/- Mice Biol Reprod, September 1, 2001; 65(3): 771 - 776. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. H. G. J. Schrans-Stassen, H. J. G. van de Kant, D. G. de Rooij, and A. M. M. van Pelt Differential Expression of c-kit in Mouse Undifferentiated and Differentiating Type A Spermatogonia Endocrinology, December 1, 1999; 140(12): 5894 - 5900. [Abstract] [Full Text]
|
|
|
|
|
|
B. Lassalle, A. Ziyyat, J. Testart, C. Finaz, and A. Lefevre Flow cytometric method to isolate round spermatids from mouse testis Hum. Reprod., February 1, 1999; 14(2): 388 - 394. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
