目的:体外分离培养大鼠胚胎胰腺间质细胞(pancreas mesenchymal cells)。方法:以孕12.5天(E12.5)的大鼠胚胎为组织来源,通过显微分离得到E12.5的胚胎胰腺,在鼠尾胶原包覆的六孔板内进行组织块培养,观察培养过程中胰芽的形态变化,间质细胞的长出,并用细胞免疫荧光的方法对间质细胞进行了鉴定。结果:显微分离得到了胚胎胰腺,为间质细胞群包绕着上皮细胞团的结构,培养第二天即发现间质细胞的长出,培养第四天时得到间质细胞群,免疫荧光鉴定为vimemin阳性的间质细胞。结论:本方法通过大鼠E12.5胚胎胰腺显微分离,体外培养出原代胚胎胰腺间质细胞,为进一步研究大鼠胰腺间质细胞的功能提供实验平台。
Objective: Isolation and culture of E12.5 rat pancreas mesenchymal cells. Methods: The pancreas was dissected by micro-dissection from the E12.5 rat embryo, then cultured in rat tail collagen coated 6-well plate. The morphologic change and mesenchymal cells' outgrowth were detected, and the cell type was characterized by cell immunofluorescence. Results: The embryonic pancreas was successfully dissected. They appeared to be a construction of epithelial cells surrounded by mesenchymal cells. The mesenchymal cells began outgrowing when cultured for 2 days and formed mesenchymal cell mass after 4 days' culture. The cell mass was characterized by immunofluorescence of vimentin, a mesenchymal cell marker. Conclusions: In this study, the E12.5 pancreata have been successfully dissected and the primary mesenchymal cells were cultured, which can provide a good platform for the study on the function of the mesenchymalcells.