小鼠胚胎腭突间充质细胞的分离与体外培养方法研究

Separation and Culture of Mouse Embryonic Palatal Mesenchymal Cells in vitro

目的改进小鼠胚胎腭突取材方法,原代培养C57BL/6J小鼠胚胎腭突间充质细胞并加以纯化。方法通过改良方法在显微镜下切取胚胎腭突,然后应用DISPASE酶消化离体腭突,纯化腭突胚胎间充质细胞,免疫荧光染色法鉴定腭突间充质细胞生物学特性。结果改良的取材方法使小鼠胚胎腭突取材更精确,操作更简单;纯化后的原代间充质细胞几乎不含上皮细胞;免疫荧光显示细胞HNK-1、S-100、波形蛋白标记阳性,角蛋白标记阴性。结论建立了一种小鼠胚胎腭突精确取材及纯化腭突外胚间充质细胞的方法,为下一步研究工作打下基础。

Objective To modify the operation of dissecting embryonic palatal shelves and purify the mouse embryonic palatal mesenchymal （EPM） cells in primary culture. Methods The embryonic palatal shelves were dissected using a surgical microscope by modified operation. Then the embryonic palatal shelves were incubated with Dispase and the isolated EPM cells were cultured. Immunofluorescence technique was used to identify the characteristics of cells. Results Embryonic palatal shelves could be dissected accurately and easily with a modified operation. The purified EPM cells contained scarcely epithelial cells. EPM cells were anti-HNK-1, S-100, vimentin positive and anti-CK negative. Conclusion A modified method for dissecting embryonic palatal shelves and purifying the EPM cells of primary culture was established.