猪肺脏巨噬细胞的分离培养

The Porcine Lung Macrophages Isolation and Culture in Vitro

提取分离猪肺泡巨噬细胞(PAM)、猪肺血管巨噬细胞(PIM)、猪肺间质巨噬细胞(IM)三种细胞并进行体外培养,采用免疫组化和流式细胞术进行细胞的异质性研究。无菌条件下通过支气管肺泡灌洗获取PAM,右心室插管经心脏行肺血管原位灌注的方法分离得到PIM,肺组织机械处理结合酶消化法(胶原酶IA)分离IM。显微镜下观察三种细胞的形态上的不同;免疫组化和流式细胞术检测细胞表型。三种巨噬细胞形态有明显的不同,PAM较大,表面粗糙,PIM和IM较小,并且细胞光滑;PAM细胞大多为CD163阳性,IM和PIM则CD86阳性较多。PAM、PIM、IM在形态和表型上存在异质性。

Extracting and separating porcine alveolar macrophages（PAM）, pulmonary intravascular macrophages（PIM）, interstitial macrophages（IM）, three types of cells in vitro, using immunohistochemical technique and flow cytometry. Aseptic conditions by bronchoalveolar lavage for PAM, right ventricular intubation via in situ perfusion line cardiac pulmonary vascular PIM, were isolated from lung tissue mechanical treatment combined with enzyme digestion method（collagenase IA） separation of IM. Immunohistochemistry and flow cytometry to detect cell phenotype and microscope different in three kinds of cell morphology. There is obvious difference in morphology between PAM, PIM and IM. PAM is bigger and rough surface, but PIM, IM are smaller and smooth surface. PAM are mostly CD163 positive cells, IM and PIM are mostly CD86 positive PAM, PIM and IM are heterogenous in morphology and phenotype.