Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
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Ovarian cell aggregate culture in teleost, marine medaka (Oryzias dancena): basic culture conditions and characterization

  • Jae Hoon, Choi (Department of Fisheries Biology, Pukyong National University) ;
  • Seung Pyo Gong (Department of Fisheries Biology, Pukyong National University)
  • Received : 2023.12.14
  • Accepted : 2024.01.18
  • Published : 2024.03.31
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Abstract

Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

Keywords

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) grant funded from the Korea government (MSIT) (No. 2019R1F1A1058145) and by the National Institute of Fisheries Science, Ministry of Oceans and Fisheries, Korea (R2024019).

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