Cryopreservation and xenotransplantation of spotted sea bass (Lateolabrax maculatus) spermatogonia

  • Anim Reprod Sci. 2026 Jul:290:108180. doi: 10.1016/j.anireprosci.2026.108180.
Jieyun Guo  1 ,  Lulu Yan  2 ,  Chao Zhao  3 ,  Bo Zhang  3 ,  Jiangtian Lin  3 ,  Lihua Qiu  4 ,  Shengkang Li  5
Affiliations
  • 1. Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou 515063, China; State Key Laboratory of Mariculture Biobreeding and Sustainable Goods (BRESG), Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China.
  • 2. College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • 3. State Key Laboratory of Mariculture Biobreeding and Sustainable Goods (BRESG), Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China; Sanya Tropical Fisheries Research Institute, Sanya, China.
  • 4. State Key Laboratory of Mariculture Biobreeding and Sustainable Goods (BRESG), Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China; Sanya Tropical Fisheries Research Institute, Sanya, China. Electronic address: [email protected].
  • 5. Guangdong Provincial Key Laboratory of Marine Biology, Shantou University, Shantou 515063, China. Electronic address: [email protected].
PMID: 41955652 DOI: 10.1016/j.anireprosci.2026.108180
Abstract

Cryopreservation of spermatogonia (SGs) is crucial for aquaculture, broodstock innovation, and the long-term conservation of aquatic genetic resources. This study developed an efficient cryopreservation protocol for spotted sea bass SGs and evaluated their post-thaw viability and transplantation potential. Initially, we characterized testicular tissue from 6-month-old male spotted sea bass, identifying various germ cell stages and strong Lmvasa expression in SGs. An optimized method for isolating and enriching SGs using Percoll density gradient centrifugation yielded a highly viable (99.6%) SG population, confirmed by morphology and germline markers (Lmvasa, Lmdnd, Lmnanos3). Subsequently, a cryopreservation protocol was optimized, with the optimal cryomedia comprising 1.3 M DMSO + 0.1 M Trehalose + 10% FBS + 35.2% extender in MEM, achieving a remarkable post-thaw SG viability of 91.83%. The transplantation potential of both fresh and 1-year cryopreserved SGs was evaluated by xenotransplantation into medaka larvae. Both fresh and 1-year cryopreserved SGs successfully colonized the genital ridges and formed germline chimeras, with 25% chimerism in fresh SGs and 20% in cryopreserved SGs, as confirmed by donor-specific Lmvasa 3'UTR PCR analysis. This study establishes a comprehensive system for SG isolation, cryopreservation, and functional assessment, providing a foundational tool for marine germplasm banking and surrogate broodstock technology.

Keywords
Germ cell transplantation; Spotted sea bass cells; Testicular cryopreservation; Xenogeneic germline chimera.
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