BACKGROUND: Pulmonary microvascular endothelial cells (PMVECs) are one of the most important endothelial cell models for microcirculation. During many training methods, simple adherence method is relatively simple, but the time-consuming and more impurity cells are the biggest obstacle for cells culture.
OBJECTIVE: To optimize the cultivation method for mouse PMVECs in vitro, and to observe the growth condition and to identify the property.
METHODS: The pulmonary tissue particles were isolated from 5-day-old C57BL/6J mice pups by scissoring peripheral lung lobes accurately under sterile conditions, and cultured with endothelial cell medium. The morphology and growing characters of the cells were observed by inverted microscope. Cultured cells were identified by immunohistochemistry and immunofluorescence staining of factor Ⅷ related antigen.
RESULTS AND CONCLUSION: A few spindle-shaped cells were found along the edge of tissue particles after cultured for 24 hours. After subculturing, cells spread even faster and took on a homogeneous cobblestone-like appearance, with the purity exceeding 98%. Morphological observations and detections of Ⅷ related antigen results conformed that these cultured cells were endothelial cells. It is a convenient and effective approach to gain a large scale of PMVECs from neonatal mice by tissue-anchoring method and endothelial cell medium.