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Isolation,culture, and characterization of primordial germ cells in Mongolian sheep
Authors:Chun Xia Liu  Wen Long Wang  Rui Yuan Zhao  Hai Tao Wang  Yi Yi Liu  Shen Yuan Wang  Huan Min Zhou
Affiliation:1. College of Life Science, Inner Mongolia Agricultural University, 306 Zhao wu da street, Hohhot, Inner Mongolia, 010018, China
2. College of Veterinary Medicine, Inner Mongolia Agricultural University, 306 Zhao wu da street, Hohhot, Inner Mongolia, 010018, China
Abstract:This study was performed to culture and preliminarily identify the primordial germ cells (PGCs) isolated from the genital ridge of the Mongolian sheep fetus. The growth characteristics of the sheep PGCs were detected in different culture systems such as culture media, resources, and state and passages of feeder cells. The obtained embryonic germ (EG) cells were identified by morphology, enzymology, and immunofluorescence. The results showed that the sheep EG cell colonies were ridgy, typically nest like, and compact, and had regular edges. Alkaline phosphatase staining reaction was weakly positive. EG cells expressed Kit, Rex-1, Nanog, and Oct-4. Immunofluorescence detection was weakly positive for Oct3/4, whereas positive for SSEA-1, SSEA-3, SSEA-4, TRA-1-61, and TRA-1-80.
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