Construction of directional differentiation model from mouse embryonic stem cells to Leydig-like cells in vitro / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 386-392, 2012.
Artigo
em Chinês
| WPRIM
| ID: wpr-336779
ABSTRACT
<p><b>OBJECTIVE</b>To construct a directional differentiation model from mouse embryonic stem cells into leydig-like cells in vitro.</p><p><b>METHODS</b>Mouse ES-D3 cells were transfected with plasmid containing steroidogenic factor 1 (SF-1) gene, then treated with RA and 8Br-cAMP, while the cells transfected with empty plasmid were used as the negative controls. The morphology of leydig-like cells differentiated from ES-D3 cells was observed with light microscopy. The expression levels of StAR, P450scc and 3β-HSD were detected by RT-PCR, Western Blot and fluorescence microscopy analysis in leydig-like cells derived from the ES cells.</p><p><b>RESULTS</b>ES-D3 cells were transfected with plasmid containing SF-1 gene successfully, and SF-1 was expressed 24 h after transfection. The SF-1-transfected ES-D3 cells were induced by RA and 8Br-cAMP to differentiate into leydig-like cells. The differentiated cells showed spindle shape with tentacles, which expressed the specific protein marker for leydig cells 3β-HSD1 and P450scc. Meanwhile, in these leydig-like cells, the expression of StAR increased compared with control group. 3β-HSD1, P450scc and StAR were not detected in negative control group.</p><p><b>CONCLUSION</b>When the ES-D3 cells are transfected with SF-1 plasmid and then treated with RA and 8Br-cAMP, the cells are able to differentiate into leydig-like cells, indicating that the model of directional differentiation of ES cells into leydig-like cells has been constructed successfully.</p>
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Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Transfecção
/
Diferenciação Celular
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Linhagem Celular
/
Biologia Celular
/
Células-Tronco Embrionárias
/
Fator Esteroidogênico 1
/
Genética
/
Células Intersticiais do Testículo
/
Metabolismo
Tipo de estudo:
Estudo prognóstico
Limite:
Animais
Idioma:
Chinês
Revista:
Journal of Zhejiang University. Medical sciences
Ano de publicação:
2012
Tipo de documento:
Artigo
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