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This Article Full Text Full Text (PDF) All Versions of this Article: 76/1/173    most recent biolreprod.106.056275v1 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 Google Scholar Google Scholar Articles by Jung, J. G. Articles by Han, J. Y. Search for Related Content PubMed PubMed Citation Articles by Jung, J. G. Articles by Han, J. Y. Agricola Articles by Jung, J. G. Articles by Han, J. Y.

Identification, Culture, and Characterization of Germline Stem Cell-Like Cells in Chicken Testes¹

Department of Food and Animal Biotechnology,³ Seoul National University, Seoul 151-921, Korea Avicore Biotechnology Institute Inc.,⁴ Gunpo, Gyeonggi-Do 435-824, Korea Research Institute for Agriculture and Life Sciences,⁵ Seoul National University, Seoul 151-921, Korea

ABSTRACT

We recently succeeded in inducing germline transmission by transferring chicken testicular cells into heterologous testes. This study was designed subsequently to identify pluripotent cells in the testicular cells, which would induce the germline transmission. Testicular cells retrieved from juvenile (4-wk-old) or adult (24-wk-old) White Leghorn (WL) chickens were stained with germ cell-specific markers anti-SSEA1, anti-SSEA3, anti-SSEA4, anti-EMA1, anti-ITGA6, and anti-ITGB1 antibodies; 2C9; and lectin-Solanum tuberosum agglutinin (STA). The percentages of the cells that were positive for each marker were within the ranges of 0.33%–0.44% and 0.029%–0.072% of the total testicular cell population in the juvenile and adult, respectively, and significant (P < 0.0002) differences were detected between the ages. When 1 x 10⁶ testicular cells were cultured in Dulbecco minimum essential medium-based medium supplemented with leukemia inhibitory factor (LIF), basic fibroblast growth factor (FGF2), and/or insulinlike growth factor 1 (IGF1), colony formation was detected only in LIF++FGF2-containing or LIF+FGF2+IGF1-containing medium during primary culture, and the supplementation of LIF+FGF2+IGF1 was the most efficient for maintaining the colony-forming cells through subculture. The established cells retrieved at the end of the primary culture or the 20th subpassage were positive for chicken germ cell-specific periodic acid-Schiff (PAS), EMA1, 2C9, SSEA1, SSEA3, SSEA4, ITGA6, and ITGB1; and lectin-STA markers (evaluated after 11th subpassage). Double staining of lectin-STA with anti-SSEA1, anti-SSEA3, anti-SSEA4, anti-ITGA6, and anti-ITGB1 also was possible. They differentiated spontaneously into embryoid bodies after being cultured in LIF-free medium. We conclude that germline stem cell-like cells are present in chicken testicular cells retrieved from both juvenile and adult testes, which can be identified with the specific markers for primordial germ cells or embryonic germ cells.

chicken, culture, cytokines, developmental biology, gamete biology, germ cell-specific marker, germline stem cell, spermatogenesis, testis

¹Supported by a graduate fellowship from Brain Korea 21 project, Republic of Korea.

Correspondence: ² Jae Yong Han, Division of Animal Genetic Engineering, School of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Korea. FAX: 822 874 4811; e-mail: jaehan{at}snu.ac.kr