肉鸡肺微血管内皮细胞的培养与鉴定

A simple and easy method for cultivation of avian pulmonary microvascular endothelial cells

选用17d鸡胚,采用组织块培养法,在无菌条件下分离培养肺微血管内皮细胞(PMVECs),60h移除组织块,结合细胞形态和免疫细胞化学对培养细胞进行鉴定。结果表明,培养24h组织块周围即有细胞爬出,48h后细胞大量增殖。形状以梭形或多边形居多,细胞扁平排列紧密,大小均匀,融合为单层后呈现出典型的鹅卵石外观。异植物血凝素(FITC-BSI)结合试验、CD31单抗和Ⅷ因子相关抗原免疫染色均呈阳性反应,证明分离培养细胞为PM-VECs。本试验提出的组织块培养法操作简单,对仪器设备要求不高,能较快且稳定地在体外培养出PMVECs,该方法的建立有助于家禽心血管疾病,如肺动脉高压、腹水和肺血管结构重构等相关疾病的深入研究。

Tissue culture method was used to （PMVECs）from 17-day embryonated eggs. cultivate pulmonary microvascular endothelial cells The cultured cells were identified by cellular mor- phology and immunocytochemistry. The results revealed that a few cells migrated out of tissue pieces at 24 h of cultured,and underwented extensive proliferation after 48 h. Under inverted mi- croscope,the confluent cells exhibited a typical cobblestone morphology. The cultured cells had characterization of binding lectin from Bandeiraea simplicifolia and positive immunocytochemi- cal staining with CD31 antibody and factor Ⅷ related antigen. This proposal provides a relative simple,reliable method of isolating PMVECs in vitro culture conditions,which can be used for study in avian cadiovascular disease such as pulmonary hypertension, ascites syndrome and pulmo- nary vascular structural remodelling.