摘要
实验应用SD雄性大鼠的外周肺组织,进行大鼠肺微血管内皮细胞(RPMVEC)的原代培养和传代培养,并从实验动物、培养基、操作条件、剪块方法、植块方法等细节方面对组织块法培养RPMVEC的方法进行探讨改进。同时,对培养细胞进行Ⅷ因子相关抗原免疫细胞化学鉴定,通过光镜和透射电镜观察细胞形态和超微结构,通过MTT观察细胞生长情况。体外培养的原代RPMVEC在光镜下呈多角形或梭形,胞核清晰,呈卵圆形。细胞单层呈典型的铺路石样排列。细胞生长状态良好、抗Ⅷ因子相关抗原染色阳性。透射电镜可见Weibel-Palade小体。改进后的组织块培养法获取RPMVEC的成功率高,污染几率低,细胞活力好,纯度高,数量大,同时保有了PMVEC的结构和功能,可用于进一步的实验研究。
To establish a simple and effective method of culturing rat Pulmonary Microvascular Endothelial Cells （ RPMVECs）, the tissues of lung outer edge of Sprague-Dawley rats to culture RPMVECs is employed. The method was improved from animal, culture medium, operation condition, tissue reduced and implanted aspects. The RPMVECs was identified by transmission electron microscope and immunofluorescence studies of factor Ⅷ. The cell rate was evaluated by MTT methods. The primary cells and subcuhured cells showed orbicular-ovate nucleus crocopy. Native RPMVECs exhibited numerous vesicles and occasional Weibel-Palade bodies along with some mitochondria and rough endoplasmic reticulum. Immunofluorescence of anti-factor Ⅷ antigen was positive. The data obtained clearly establish that the culture of primary RPMVECs by improved methods of implanting tissue pieces is effective and successful. The cells and function as well, which can obtained by our methods are good in quantity and purity and maintain its structure be applied into further experiment.
出处
《科学技术与工程》
2008年第5期1160-1164,共5页
Science Technology and Engineering
关键词
大鼠肺微血管内皮细胞
培养方法
组织
rat pulmonary microvascular endothelial cells culture methods strueture
