HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Takeuchi, Y. Articles by Takeuchi, T. Search for Related Content PubMed PubMed Citation Articles by Takeuchi, Y. Articles by Takeuchi, T. Agricola Articles by Takeuchi, Y. Articles by Takeuchi, T.

Mass Isolation of Primordial Germ Cells from Transgenic Rainbow Trout Carrying the Green Fluorescent Protein Gene Driven by the vasa Gene Promoter

^a Department of Aquatic Biosciences, Tokyo University of Fisheries, Minato-ku, Tokyo 108-8477, Japan

Mass isolation of live primordial germ cells (PGCs) was demonstrated for the first time in ectothermal vertebrates. To establish a stem cell-mediated gene transfer system in fish, a stem cell line that retains the ability to develop into gametes is necessary. PGCs are well suited for use as the initial material for such a stem cell line. We established transgenic rainbow trout (Oncorhynchus mykiss) strains carrying the green fluorescent protein (GFP) gene driven by a rainbow trout vasa-like gene (RtVLG) promoter/enhancer. Because GFP expression was specific to the PGCs, PGCs were successfully visualized in all developmental stages examined. Isolated genital ridges containing GFP-labeled PGCs were enzymatically dissociated. To isolate PGCs from the complex pools of dissociated genital ridges, GFP-labeled cells were sorted by flow cytometry. The sorted GFP-positive cells were large and round with a large nucleus, typical characters of PGC morphology. The expression of RtVLG was detected only in the GFP-positive cell population, confirming that these cells were PGCs. This simple and efficient technique to purify a large number of viable PGCs opens the way for establishing a stem cell line, which can differentiate into the germline. The purified PGCs would also be a novel tool for cellular and molecular study of vertebrate germline stem cells.

¹ Correspondence: Goro Yoshizaki, Department of Aquatic Biosciences, Tokyo University of Fisheries, 4–5–7 Konan, Minato-ku, Tokyo 108–8477, Japan. FAX: 81 3 5463 0558; goro{at}tokyo-u-fish.ac.jp

This article has been cited by other articles:

				A. Yano, K. Suzuki, and G. Yoshizaki Flow-Cytometric Isolation of Testicular Germ Cells from Rainbow Trout (Oncorhynchus mykiss) Carrying the Green Fluorescent Protein Gene Driven by Trout vasa Regulatory Regions Biol Reprod, January 1, 2008; 78(1): 151 - 158. [Abstract] [Full Text] [PDF]

				T. Okutsu, K. Suzuki, Y. Takeuchi, T. Takeuchi, and G. Yoshizaki Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish PNAS, February 21, 2006; 103(8): 2725 - 2729. [Abstract] [Full Text] [PDF]

				G. Yoshizaki, Y. Tago, Y. Takeuchi, E. Sawatari, T. Kobayashi, and T. Takeuchi Green Fluorescent Protein Labeling of Primordial Germ Cells Using a Nontransgenic Method and Its Application for Germ Cell Transplantation in Salmonidae Biol Reprod, July 1, 2005; 73(1): 88 - 93. [Abstract] [Full Text] [PDF]

				Y. Takeuchi, G. Yoshizaki, and T. Takeuchi Generation of Live Fry from Intraperitoneally Transplanted Primordial Germ Cells in Rainbow Trout Biol Reprod, October 1, 2003; 69(4): 1142 - 1149. [Abstract] [Full Text] [PDF]