大鼠骨髓内皮细胞的分离培养及其形态特征

目的：分离培养大鼠骨髓内皮并进行生物学鉴定。方法：应用Peroll细胞分离介质（密度1．04－1．06g/ml)，将SD大鼠长骨骨髓经非连续梯度离心后收集位于中层的细胞进行培养，观察细胞生长状况，绘制细胞生长曲线并以透射电镜及Ⅷ因子免疫组化方法对培养细胞进行鉴定。结果：细胞圆型单层融合贴壁生长，平均直径15μm，单个核并位中央，群体倍增时间约为37h；Ⅷ因子免疫组化染色阳性；透射电镜显示细胞内具有特征性的W－P小体。结论：采用非连续梯度离心沉降法可获得高纯度的骨髓内皮细胞，该细胞具有与血管内皮细胞共同的特征。

Culture and characterization of rat bone marrow endothelial cells in vitro

Objective: To isolate, culture and observe the biological characteristics of endothelial cells derived from rat bone marrow. Methods: Marrow cell suspension was prepared by discontinuous gradient centrifugation on Percoll (densities 1.04～1.06 g/ml). Endothelial cells were concentrated in the middle layer and subjected to culture in vitro with Gibco's vascular endothelial cell culture medium. The cells were identified by transmission electron microscopy and immunohistochemical staining with factor Ⅷ antigen. The cells was observed under inverted microscope every day and cell growth was studied with cell counting. Results: The cells uniformly had a rounded shape with an average cell diameter of 15 μm. The population doubling time of the cells was 37 h. The cells were factor-Ⅷ positive. Undertransmission electron microscope, the cells displayed many fingerlike microvilli and numerous lysosomes, mitochondria, a few rough endoplasmid reticulum and Weibel-Palade bodies. Conclusions: It seems that density-gradient centrifugation sedimentation is an effective approach to isolate rat bone marrow endothelial cells. The characteristics of the rat bone marrow endothelial cells in vitro are consistent with those of vascular endothelial cells.