Preservation of endemic fish using germ stem and iPS cells

2014 Fiscal Year Final Research Report

• Project/Area Number: 23651248
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Resource conservation science
• Research Institution: Ritsumeikan University
• Principal Investigator: TAKADA tatsuyuki 立命館大学, 薬学部, 教授 (90206756)
• Co-Investigator(Renkei-kenkyūsha): SAKAI Noriyoshi 国立遺伝学研究所, 系統生物研究センター, 准教授 (50202081) ; INOUE Hirokazu 滋賀医科大学, 医学部, 准教授 (30176440)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Keywords: 精原細胞 / in vitro 精子分化 / 生殖細胞 / 細胞株樹立 / ホンモロコ / 固有種保全 / 環境ホルモン / 琵琶湖

Outline of Final Research Achievements

Many endemic fish species inhabit in Lake Biwa. The number of such species, however, is rapidly decreasing in recent decades. We tried to apply stem cell biology for preservation of endemic fish. We succeeded to develop culture method and establish a total of 19 types of cell line from testis, ovary and fertilized embryo in endemic fish Honmoroko (Gnathopogon caerulescens). These cell line were useful to validate hormone response and endocrine disrupting chemicals, and we found that nonylphenol inhibited androgenic effect in G. caerulescence. We also developed a method to analyze somatic and germ cells of various developmental stages quantitatively using flow cytometry. Furthermore, we succeeded to differentiate fertilization competent sperm from cryopreserved spermatogonia in vitro. Although generation of iPS cells requires further study, we established a new strategy for preservation of endemic fish combining in vitro formation of gametes and cryopreservation of germ stem cells.

Free Research Field

幹細胞生物学