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KMID : 1161520150190030168
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Animal Cells and Systems
2015 Volume.19 No. 3 p.168 ~ p.174
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Embryoid bodies formation from chicken primordial germ cells
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Xiong Hui
Pu Yabin
Hu Qingyun
Shan Zhiqiang
Hu Pengfei
Guan Weijun
Ma Yuehui
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Abstract
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Primordial germ cells (PGCs) were demonstrated to be multipotential because of their differentiation ability in all embryonic lineages and because the pluripotential nature of PGCs caters for recent researches on stem cells. PGCs were cultured in suspension to form embryoid bodies (EBs). The characterization of PGCs and EBs was assessed by immunofluorescence technique, reverse transcription-polymerase chain reaction (RT-PCR) and paraffin section for Haematoxylin and Eosin (HE) stain. We established a stable chicken PGC line in vitro and prepared masses of EBs from PGCs. We also demonstrated that PGCs expressed stage-specific and stem cell-specific surface makers: SSEA-1, SSEA-3, Oct4, and Sox2. EBs expressed specific genes from three germ layers: gene AFP from entoderm, gene GATA6 from mesoderm, and gene Sox3 from ectoderm. The HE staining illustrated that EBs developed different cell types; relatively larger EBs (440 μm in diameter) were obtained, which contract rhythmically. We concluded that chicken PGCs were suitable for the formation of EBs. Abundant larger size EBs could be prepared in an effective way with our protocol, which could construct a good animal model and provide a useful clinical platform in studying human embryonic development and cell transplantation field.
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KEYWORD
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embryoid bodies, primordial germ cells, differentiation
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