Characterization and In Vitro Differentiation of Korean Ring-Necked Pheasant (Phasianus colchicus) Male Germ Cells

• Published in: Journal of animal reproduction & biotechnology (Online) pp. 351 - 359
• Main Authors: 정동기, 니레시사르마, 김종현, 오성종
• Format: Journal Article
• Language: English
• Published: 사단법인 한국동물생명공학회 01.12.2014
• Subjects: 축산학
• ISSN: 2671-4639; 2671-4663

Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongatedsperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the differentadherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cellsexamined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were inducedfor 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulatedfactors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptionalmarker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatidbasic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1CDNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successfulin vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potentialfor the conservation of endangered birds and treatment of male infertility. KCI Citation Count: 0