Challenge to establish the immortalized shrimp cell line

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K20299
• Project/Area Number (Other): 17H06249 (2017-2019)
• Research Category: Grant-in-Aid for Challenging Research (Pioneering)
• Allocation Type: Multi-year Fund (2020); Single-year Grants (2017-2019)
• Research Field: Forestry and Forest Products Science, Applied aquatic science, and related fields
• Research Institution: University of Miyazaki
• Principal Investigator: Sakai Masahiro 宮崎大学, 農学部, 教授 (20178536)
• Co-Investigator(Kenkyū-buntansha): 引間 順一 宮崎大学, 農学部, 教授 (70708130) ; 稲田 真理 国立研究開発法人水産研究・教育機構, 水産技術研究所(南勢), 研究員 (50723558)
• Project Period (FY): 2017-06-30 – 2022-03-31
• Keywords: エビ類 / 細胞培養 / 細胞増殖因子 / ウイルス疾病

Outline of Final Research Achievements

In the present study, kuruma shrimp (Marsupenaeus japonicus) lymphoid organs were used for primary cell culture. First, the osmotic pressure of the cell culture medium was adjusted it to 780 mmol/kg. Thereafter, we determined the change in the percentage of live to dead cells after cell seeding. At two time points, days 3-4 and 6-7, the proportion of dead cells increased considerably. Next, transcriptomic analysis using RNA sequencing (RNA-seq) and quantitative real-time PCR (qPCR) was conducted on cell samples on days 1, 3, 4, and 6 after seeding. Genes downregulated on day 4 included MjPDGFRA and MjVEGF3. Consistently, relative quantification by qPCR revealed a significant decrease in the expression levels of MjVEGF3 and its receptor, MjVEGFR genes. These results suggest that a decrease in the expression levels of these genes may be related to cell growth and proliferation in primary cultures of shrimp lymphoid organ cells.

Free Research Field

魚介類疾病

Academic Significance and Societal Importance of the Research Achievements

本研究課題では、細胞の機能解析やウイルス研究を行うにあたり株化細胞は必須なツールであるエビ類の長期培養可能な細胞株の樹立が試みた。また、クルマエビ細胞の長期培養に最適なサプリメント分子の特定ができたことは、長期間培養可能なクルマエビ株化細胞を樹立するために今後大いに役立つと考えられる。