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Differentiation and characteristics of the enhanced green fluorescent protein gene transgenic goat neural stem cells cultured in attached and non‐attached plates
Author(s) -
Zheng YueMao,
Dang YongHui,
Qiu Shuang,
Qi YingPei,
Xu YongPing,
Sai WuJiaFu
Publication year - 2011
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20100899
Subject(s) - neurosphere , neural stem cell , stem cell , homeobox protein nanog , nestin , sox2 , biology , transfection , microbiology and biotechnology , embryonic stem cell , green fluorescent protein , adult stem cell , hes1 , cell culture , gene , induced pluripotent stem cell , notch signaling pathway , biochemistry , genetics , signal transduction
The aims of this study were (i) to determine whether NSCs (neural stem cells) could be isolated from the brain of embryonic day 98 fetal goat, (ii) to determine if these stem cells have the capability of multipotent differentiation following transfection with a reporter gene, EGFP (enhanced green fluorescent protein) and (iii) to study the characteristics of the stem cells cultured in attached and non‐attached plates. NSCs were isolated from embryonic day 98 fetal goat brain, transfected with EGFP gene using lipofection, and subcultured in attached and non‐attached plates respectively. The transgenic stem cells were induced to differentiate into osteogenic and endothelial cells in vitro respectively. Markers associated with undifferentiated NSCs and their differentiated cells were tested by RT—PCR (reverse transcription—PCR). The results demonstrated that stem cells could be isolated from embryonic day 98 fetal goat brain, and EGFP gene could be transfected into the cells. The transgenic NSCs were capable of self‐renewal, a defining property of stem cells, and were grown as free‐floating neurospheres in non‐attached plates. When the neurospheres were transferred and cultured in attached plates, cells migrate from the neurospheres and are grown as spindle cells. The stem cells were grown as quasi‐circular cells when the single stem cells were cultured in attached plates. Both the NSCs cultured in non‐attached and attached plates could express Hes1 (hairy and enhancer of split 1), Oct4 (octamer‐binding protein 4), Nanog, Sox2 [SRY (sex‐determining region Y)‐box 2] and Nestin, while following differentiation cells expressed markers for osteogenic cells (Osteocalcin + and Osteonectin + ) and endothelium (CD34 + and eNOS + ). The results demonstrated that the goat EGFP gene transgenic NSCs have the capability of multipotent differentiation, which means that the transgenic NSCs may be useful in cell transplantation studies in future.