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.

질경이를 공시하여 펄라이트, 펄라이트와 피트모스 1:1 혼합, 입상암면 및 피트모스 등 4종의 고형배지경에서 배지에 따른 생장반응과 무기성분 함량을 비교 분석하였다. 배지종류별 생육반응은 초장, 경경, 엽수, 최대근장, 지상부와 지하부의 생체중 및 건물중 등 전반적인 초기생장이 펄라이트와 피트모스 혼합배지〉피트모스 배지〉입상암면 배지〉펄라이트 배지 처리구 등의 순인 것으로 나타났다. 식물체내 Ca와 Mg함량은 피트모스 배지에서, Mg와 Na 함량은 입상암면 배지에서 가장 높은 것으로 나타났다. 식물체내 P2O5 함량은 식물 생장과 비례적으로 증가하여 펄라이트와 피트모스 1:1 혼합 배지 처리구와 피트모스 배지 처리구에서 높게 나타났다.