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Establishment of the Long‐Term In Vitro Culture System for Chicken Primordial Germ Cells
Author(s) -
Shiue YL,
Tailiu JJ,
Liou JF,
Lu HT,
Tai C,
Shiau JW,
Chen LR
Publication year - 2009
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/j.1439-0531.2007.00990.x
Subject(s) - biology , in vitro , embryonic stem cell , microbiology and biotechnology , embryo , haematopoiesis , andrology , gonadal ridge , somatic cell , immunology , embryogenesis , stem cell , genetics , medicine , gene
Contents The objective of this study was to establish the long‐term in vitro culture system for chicken gonadal primordial germ cells (gPGCs). Primitive gonads collected from 5.5‐day‐old chicken embryos were dissociated and explanted onto plates pre‐coated with 0.1% gelatin. Each of the four different conditioned media from proliferating and mitotically inactivated chicken embryonic fibroblast (CEF) cells and murine embryonic fibroblasts (STO cells, CRL‐1053, ATCC, USA), respectively, was supplemented with growth factors and used to support the growth of gPGCs. The result showed that all the conditioned media could promote the growth and colony formation of gPGCs in vitro , in particular the medium conditioned by inactivated CEF cells. The gPGC‐derived colonies maintained in inactivated CEF cells‐conditioned medium up to 281 days were positively stained by periodic acid Schiff reaction and antibodies specific to anti‐SSEA‐1, SSEA‐3, SSEA‐4, integrin α6 and integrin β1. Their capacities of migration via vascular system and taking up residence in the primary gonadal ridge were further demonstrated by transferring to the dorsal aorta of stage 17 recipient embryos. These results suggested that our culture system is able to maintain chicken gPGCs for long‐term in vitro culture without losing their capacity to express pluripotent markers and to integrate into the gonads.