Fish Physiology and Biochemistry

, Volume 44, Issue 2, pp 717–733 | Cite as

Testicular germ line cell identification, isolation, and transplantation in two North American catfish species

  • Mei Shang
  • Baofeng Su
  • Dayan A. Perera
  • Ahmed Alsaqufi
  • Elizabeth A. Lipke
  • Sehriban Cek
  • David A. Dunn
  • Zhenkui Qin
  • Eric Peatman
  • Rex A. Dunham


Our aim was to transplant blue catfish germ line stem cells into blastulae of triploid channel catfish embryos to produce interspecific xenogenic catfish. The morphological structure of the gonads of blue catfish (Ictalurus furcatus) in ~ 90- to 100-day-old juveniles, two-year-old juveniles, and mature adults was studied histologically. Both oogonia (12–15 μm, diameter with distinct nucleus 7–8 μm diameter) and spermatogonia (12–15 μm, with distinct nucleus 6–7.5 μm diameter) were found in all ages of fish. The percentage of germ line stem cells was higher in younger blue catfish of both sexes. After the testicular tissue was trypsinized, a discontinuous density gradient centrifugation was performed using 70, 45, and 35% Percoll to enrich the percentage of spermatogonial stem cells (SSCs). Four distinct cell bands were generated after the centrifugation. It was estimated that 50% of the total cells in the top band were type A spermatogonia (diameter 12–15 μm) and type B spermatogonia (diameter 10–11 μm). Germ cells were confirmed with expression of vasa. Blastula-stage embryos of channel catfish (I. punctatus) were injected with freshly dissociated blue catfish testicular germ cells as donor cells for transplantation. Seventeen days after the transplantation, 33.3% of the triploid channel catfish fry were determined to be xenogenic catfish. This transplantation technique was efficient, and these xenogenic channel catfish need to be grown to maturity to verify their reproductive capacity and to verify that for the first time SSCs injected into blastulae were able to migrate to the genital ridge and colonize. These results open the possibility of artificially producing xenogenic channel catfish males that can produce blue catfish sperm and mate with normal channel catfish females naturally. The progeny would be all C × B hybrid catfish, and the efficiency of hybrid catfish production could be improved tremendously in the catfish industry.


Blue catfish Germ line stem cell Spermatogonial stem cell Density gradient centrifugation Germ line transplantation 


Funding information

This work was partially funded by the USDA-Biotechnology Risk Assessment Program (grant no.: 2009-33522-05774), the Alabama Agricultural Experiment Station (80-009), and the Office of the Vice President for Research, Auburn University (OVPR-AU-IGP).

Compliance with ethical standards

All the procedures involving the handling of fish during this study were approved by the Auburn University Institutional Animal Care and Use Committee (PRN-2013-2224).

Supplementary material

10695_2018_467_MOESM1_ESM.docx (15 kb)
Supplementary figure 1 (DOCX 14kb).


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Mei Shang
    • 1
    • 2
  • Baofeng Su
    • 1
    • 2
  • Dayan A. Perera
    • 1
    • 3
  • Ahmed Alsaqufi
    • 1
    • 4
  • Elizabeth A. Lipke
    • 5
  • Sehriban Cek
    • 6
  • David A. Dunn
    • 5
    • 7
  • Zhenkui Qin
    • 1
  • Eric Peatman
    • 1
  • Rex A. Dunham
    • 1
  1. 1.School of Fisheries, Aquaculture, and Aquatic SciencesAuburn UniversityAuburnUSA
  2. 2.Key Laboratory of Freshwater Aquatic Biotechnology and Genetic Breeding, Ministry of AgricultureHeilongjiang Fisheries Research Institute, Chinese Academy of Fishery SciencesHarbinChina
  3. 3.School of Agriculture, Fisheries and Human SciencesUniversity of Arkansas at Pine BluffPine BluffUSA
  4. 4.Department of Aquaculture and Animal ProductionKing Faisal UniversityHofufKingdom of Saudi Arabia
  5. 5.Department of Chemical EngineeringAuburn UniversityAuburnUSA
  6. 6.Faculty of Marine Sciences and TechnologyIskenderun Technical Universityİskenderun/HatayTurkey
  7. 7.Department of Biological SciencesState University of New York at OswegoOswegoUSA

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