Abstract

Spermatogenesis is a continuous and dynamic developmental process, in which a single diploid spermatogonial stem cell (SSC) proliferates and differentiates to form a mature spermatozoon. Herein, we summarize the accumulated knowledge of SSCs and their distribution in the testes of teleosts. We also reviewed the primary endocrine and paracrine influence on spermatogonium self-renewal vs. differentiation in fish. To provide insight into techniques and research related to SSCs, we review available protocols and advances in enriching undifferentiated spermatogonia based on their unique physiochemical and biochemical properties, such as size, density, and differential expression of specific surface markers. We summarize in vitro germ cell culture conditions developed to maintain proliferation and survival of spermatogonia in selected fish species. In traditional culture systems, sera and feeder cells were considered to be essential for SSC self-renewal, in contrast to recently developed systems with well-defined media and growth factors to induce either SSC self-renewal or differentiation in long-term cultures. The establishment of a germ cell culture contributes to efficient SSC propagation in rare, endangered, or commercially cultured fish species for use in biotechnological manipulation, such as cryopreservation and transplantation. Finally, we discuss organ culture and three-dimensional models for in vitro investigation of fish spermatogenesis.

Highlights

  • Spermatogenesis is a complex and orderly developmental process in which a single diploid spermatogonial stem cell (SSC) proliferates and differentiates to form mature spermatozoa

  • Spermatogenesis depends on the activity of the SSC, which can both self-renew to produce more stem cells or differentiate into daughter cells committed to spermatogenesis [1,2,3,4,5]

  • The proper balance between SSC self-renewal and differentiation is essential to assure the continuous homeostasis of spermatogenesis

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Summary

Overview of Fish Germ Cell Biology

Spermatogenesis is a complex and orderly developmental process in which a single diploid spermatogonial stem cell (SSC) proliferates and differentiates to form mature spermatozoa. With respect to whether spermatogonia display a preferential distribution within the fish testes, studies of zebrafish [22] and Astyanax altiparane [45] have shown that both Aund* and Aund spermatogonia are located near the interstitial compartment, in contact with androgen-producing Leydig cells and blood vessels (Figure 4A–C). These observations suggest that the androgens; growth factors; and vascular supplies of oxygen, nutrients, and hormones may play essential roles in SSC maintenance and self-renewal vs differentiation in the fish testis niche [10,21].

Endocrine and Paracrine Regulation
Enzymatic Digestion
Germ Cell Purification
Density Gradient Centrifugation
Differential Plating
Flow Cytometric and Magnetically-Activated Cell-Sorting
Future Trends
Serum in Germ Cell Cultures
Feeder Cells and Growth Factors
Organ Cultures
Three-Dimensional Cultures

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