Spermatogenesis In Vitro: Searching for In Vivo Correlates

  • Abraham L. Kierszenbaum
  • Munir Abdullah
  • Hiroshi Ueda
  • Laura L. Tres
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 219)


The development of methodology for the study of spermatogenesis in vitro has been a major task that remained elusive (Wolff and Haffen, 1965; Steinberger, 1975). Significant advances have been reported in the culture of meiotic cells of lilaceous plants (Hotta et al., 1966), and spermatogenic cells of Xenopus (Risley, 1983) and Drosophila (Liebrich, 1981). Attempts to maintain isolated spermatogenic cells viable for long periods of time (more than 48 h) have been relatively unsuccessful. In fact, the seminiferous epithelium contains spermatogenic cells that maintain spatial and functional relationships with Sertoli cells, and this cell-cell interaction is likely to play an important role in spermatogenesis in vivo and in vitro.


Sertoli Cell Seminiferous Tubule Spermatogenic Cell Seminiferous Epithelium Pachytene Spermatocyte 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Abdullah, M, Crowell, JA, Tres, LL, Kierszenbaum, AL, 1986. Fetuin: A serum component associated with rat Sertoli and spermatogenic cells in coculture. J Cell Physiol 127:463–72PubMedCrossRefGoogle Scholar
  2. Borland, K, Mita, M, Oppenheimer, CL, Blinderman, LA, Massague, J, Hall, PH, Czech, MP, 1984. The actions of insulin-like growth factors I and II on cultured Sertoli cells. Endocrinology 114:240–46PubMedCrossRefGoogle Scholar
  3. Clermont, Y, 1972. Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal. Physiol Rev 52:198–236PubMedGoogle Scholar
  4. Clermont, Y, Perey, R, 1957. Quantitative study of the cell population of the seminiferous tubules in immature rats. Am J Anat 100:241–67PubMedCrossRefGoogle Scholar
  5. DePhilip, RM, Kierszenbaum, AL, 1982. Hormonal regulation of protein synthesis, secretion, and phosphorylation in cultured Sertoli cells. Proc Natl Acad Sci USA 79:6551–55PubMedCrossRefGoogle Scholar
  6. DePhilip, RM, Tres, LL, Kierszenbaum, AL, 1982. Stage-specific protein synthesis during rat spermatogenesis. Exp Cell Res 142:489–94PubMedCrossRefGoogle Scholar
  7. Dym, M, Fawcett, DW, 1970. The blood-testis barrier in the rat and the physiological compartmentalization of the seminiferous epithelium. Biol Reprod 3:308–26PubMedGoogle Scholar
  8. Fakunding, JL, Means, AR, 1977. Characterization and follicle stimulating hormone activation of Sertoli cell cyclic AMP-dependent protein kinases. Endocrinology 101:1358–68PubMedCrossRefGoogle Scholar
  9. Feldman, SR, Gonias, SL, Ney, KA, Pratt, CW, Pizzo, SV, 1984. Identification of “embryonin” as bovine α2 -macroglobulin. J Biol Chem 259:4458–62PubMedGoogle Scholar
  10. Feldman, M, Lea, OA, Petrusz, P, Tres, LL, Kierszenbaum, AL, French, FS, 1981. Androgen binding proteins: purification from rat epididymis, characterization, and immunocytochemical localization. J Biol Chem 256: 5170–5175PubMedGoogle Scholar
  11. Feig, L, Klagsbrun, GT, Bellvé, AR, 1983. Mitogenic polypeptide from the mammalian seminiferous epithelium: biochemical characterization and partial purification. J Cell Biol 97:1435–43PubMedCrossRefGoogle Scholar
  12. Heindel, JJ, Steinberger, A, Strada, SJ, 1981. Identification and characterization of a β -adrenergic receptor in rat Sertoli cells. Mol Cell Endocrinol 22:349–58PubMedCrossRefGoogle Scholar
  13. Hotta, Y, Ito, M, Stern, H, 1966. Synthesis of DNA during meiosis. Proc Natl Acad Sci USA 56:1184–91PubMedCrossRefGoogle Scholar
  14. Huggenvik, JI, Idzerda, RL, Haywood, L, Lee, DC, McKnight, GS, Griswold, MD, 1987. Transferrin messenger ribonucleic acid: molecular cloning and hormonal regulation in rat Sertoli cells. Endocrinology 120:332–40PubMedCrossRefGoogle Scholar
  15. Kierszenbaum, AL, Crowell, JA, Shabanowitz, RB, DePhilip, RM, Tres, LL, 1986a. Protein secretory patterns of rat Sertoli and peritubular cells are influenced by culture conditions. Biol Reprod 35:239–51PubMedCrossRefGoogle Scholar
  16. Kierszenbaum, AL, Crowell, JA, Shabanowitz, RB, Smith, EP, Spruill, WA, Tres, LL, 1986b. A monoclonal antibody recognizes a form of intermediate filament protein in rat Sertoli cells that is not present in seminiferous peritubular cells. Biol Reprod 35:227–38PubMedCrossRefGoogle Scholar
  17. Kierszenbaum, AL, Feldman, M, Lea, O, Spruill, WA, Tres, LL, Petrusz, P, French, FS, 1980. Localization of androgen-binding protein in proliferating Sertoli cells in culture. Proc Natl Acad Sci USA 77:5322–26PubMedCrossRefGoogle Scholar
  18. Kierszenbaum, AL, Spruill, WA, White, MG, Tres, LL. Perkins, JP, 1985. Rat Sertoli cells acquire a β-adrenergic response during primary culture. Proc Natl Acad Sci USA 82:2049–53PubMedCrossRefGoogle Scholar
  19. Kierszenbaum, AL, Tres, LL, 1975. Structural and transcriptional features of the mouse spermatid genome. J Cell Biol 65:258–70PubMedCrossRefGoogle Scholar
  20. Kierszenbaum, AL, Tres, LL, 1981. The structural and functional cycle of Sertoli cells in culture. In: Jagiello, G, Vogel, HJ (eds.), Bioregulators of Reproduction. New York: Academic Press, pp. 207–28Google Scholar
  21. Kierszenbaum, AL, Tres, LL, 1987. An automated perifusion system for the study of rat spermatogenesis in vitro. Ann NY Acad Sci (in press)Google Scholar
  22. Kissinger, C, Skinner, MK, Griswold, MD, 1982. Analysis of Sertoli cell secreted proteins by two-dimensional gel electrophoresis. Biol Reprod 27:233–240PubMedCrossRefGoogle Scholar
  23. Knorr, DW, Vanha-Perttula, T, Lipsett, MB, 1970. Structure and function of rat testis through pubescence. Endocrinology 86:1298–304PubMedCrossRefGoogle Scholar
  24. Leblond, CP, Clermont, Y, 1952. Definition of the stages of the cycle of the seminiferous epithelium in the rat. Ann NY Acad Sci 55:548–73PubMedCrossRefGoogle Scholar
  25. Lee, NT, Chae, CB, Kierszenbaum, AL, 1986. Contrasting levels of transferring gene activity in cultured rat Sertoli cells and intact seminiferous tubules. Proc Natl Acad Sci USA 83:8177–81PubMedCrossRefGoogle Scholar
  26. Liebrich, W, 1981. In vitro spermatogenesis in Drosophila. I. Development of isolated spermatocyte cysts from wild-type D. hydei. Cell Tissue Res 220:251–62PubMedCrossRefGoogle Scholar
  27. Libby, P, Raines, EW, Cullinane, PM, Ross, R, 1985. Analysis of the mitogenic effect of fetuin preparations on arterial smooth muscle cells: The role of contaminant platelet-derived growth factor. J Cell Physiol 125:357–66PubMedCrossRefGoogle Scholar
  28. Means, AR, Fakunding, JL, Tindall, DJ, 1976. Follicle stimulating hormone regulation of protein kinase activity and protein synthesis in testis. Biol Reprod 14:54–63PubMedCrossRefGoogle Scholar
  29. Nagy, F, 1972. Cell division kinetics and DNA synthesis in the immature Sertoli cells of the rat testis. J Reprod Fertil 28:389–95PubMedCrossRefGoogle Scholar
  30. Orth, JM, 1982. Proliferation of Sertoli cells in fetal and postnatal rats: A quantitative autoradiographic study. Anat Rec 203:485–92PubMedCrossRefGoogle Scholar
  31. Perey, B, Clermont, Y, Leblond, CP, 1961. The wave of the seminiferous epithelium in the rat. Am J Anat 108:47–77CrossRefGoogle Scholar
  32. Risley, MS, 1983. Spermatogenic cell differentiation in vitro. Gamete Res 4:331–46CrossRefGoogle Scholar
  33. Salomon, DS, Bano, M, Smith, KB, Kidwell, WR, 1982. Isolation and characterization of a growth factor (embryonin) from bovine fetuin which resembles α2-macroglobulin. J Biol Chem 257: 14093–101PubMedGoogle Scholar
  34. Setchell, BP, 1967. The blood-testicular fluid barrier in sheep. J Physiol 189:63–65Google Scholar
  35. Shabanowitz, RB, Kierszenbaum, AL, 1986. Newly synthesized proteins in seminiferous intertubular and intratubular compartments of the rat testis. Biol Reprod 35:179–90PubMedCrossRefGoogle Scholar
  36. Shabanowitz, RB, DePhilip, RM, Crowell, JA, Tres, LL, Kierszenbaum, AL, 1986. Temporal appearance and cyclic behavior of Sertoli cell-specific secretory proteins during the development of the rat seminiferous tubule. Biol Reprod 35:745–60PubMedCrossRefGoogle Scholar
  37. Smith, EP, Svoboda, ME, Van Wyk, JJ, Kierszenbaum, AL, Tres, LL, 1987. Partial characterization of a somatomedin-like peptide from the medium of cultured rat Sertoli cells. Endocrinology 120:186–93PubMedCrossRefGoogle Scholar
  38. Spruill, WA, Steiner, AL, Tres, LL, Kierszenbaum, AL, 1983a. Follicle-stimulating hormone-dependent phosphorylation of vimentin in cultures of rat Sertoli cells. Proc Natl Acad Sci USA 80:993–97PubMedCrossRefGoogle Scholar
  39. Spruill, WA, Zysk, JR, Tres, LL, Kierszenbaum, AL, 1983b. Calcium/calmodulin-dependent phosphoylation of vimentin in rat Sertoli cells. Proc Natl Acad Sci USA 80:760–64PubMedCrossRefGoogle Scholar
  40. Steinberger, E, Steinerger, A, 1970. Replication patterns of Sertoli cells in maturing rats testis in vivo and in organ culture. Biol Reprod 18: 329–45Google Scholar
  41. Sylvester, SR, Skinner, MK, Griswold, MD, 1984. A sulfated glycoprotein synthesized by Sertoli cells and by epidydimal cells is a component of the sperm membrane. Biol Reprod 31:1087–1101PubMedCrossRefGoogle Scholar
  42. Tres, LL, Kierszenbaum, AL, 1983. Viability of rat spermatogenic cells in vitro is facilitated by their coculture with Sertoli cells in serum-free hormone-supplemented medium. Proc Natl Acad Sci USA 80:3377–81PubMedCrossRefGoogle Scholar
  43. Tres, LL, Kierszenbaum, AL, 1986. Effect of substrate and medium composition on rat Sertoli-spermatogenic cell cocultures maintained in an automated cell perifusion system. IV Decennial Tissue Culture Association Review Conference (Hershey, PA), p. 24 (abstract)Google Scholar
  44. Tres, LL, Smith, EP, Van Wyk, JJ, Kierszenbaum, AL, 1986. Immunoreactive sites and accumulation of somatomedin-C in rat Sertoli-spermatogenic cell co-cultures. Exp Cell Res 162:33–50PubMedCrossRefGoogle Scholar
  45. Toews, ML, Perkins, JP, 1984. Agonist-induced changes in β-adrenergic receptors on intact cells. J Biol Chem 259:2227–35PubMedGoogle Scholar
  46. Tung, PS, Skinner, MK, Fritz, IB, 1984. Fibronectin synthesis is a marker for peritubular cell contaminants in Sertoli cell-enriched cultures. Biol Reprod 30:199–211PubMedCrossRefGoogle Scholar
  47. Vitale, R, Fawcett, DW, Dym, M, 1973. The normal development of the blood testis barrier and the effects of clomiphene and estrogen treatment. Anat Rec 176:333–44CrossRefGoogle Scholar
  48. Wolff, E, Haffen, K, 1965. Germ cells and gonads. In: Willmer, EN (ed.), Cells and Tissues in Culture, Vol 2. New York: Academic Press, pp. 697–743Google Scholar
  49. Wright, WW, Musto, NA, Mather, JP, Bardin, CW, 1981. Sertoli cells secrete both testis-specific and serum proteins. Proc Natl Acad Sci USA 78:7565–69PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Abraham L. Kierszenbaum
    • 1
  • Munir Abdullah
    • 1
  • Hiroshi Ueda
    • 1
  • Laura L. Tres
    • 1
  1. 1.Departments of Anatomy and PediatricsUniversity of North CarolinaChapel HillUSA

Personalised recommendations