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目的建立人卵巢癌微血管内皮细胞(ovarian carcinoma-derived microvascular endothelial cells,ODMECs)免疫磁珠分选体系并观察ODMECs体外二维管腔样结构(tubule-like structure,TLS)的形成。方法利用结合有CD31抗体的MACS MicroBeads免疫磁珠分选系统分离纯化ODMECs。通过形态学、生化和表型特征对所获得的ODMECs进行鉴定;并检测其摄取乙酰化低密度脂蛋白(acetylated-low-density lipoprotein,DIL-acLDL)和结合凝集素Ⅰ(UEA-Ⅰ)的能力,观察其在体外培养体系中二维管腔样结构的形成。结果所获ODMECs流式分析细胞纯度可达99.6%,具有卵石征、接触抑制,表达CD31、CD105、FⅧ-Rag;具有结合UEA-Ⅰ和摄取DIL-acLDL的能力;培养过程中观测到ODMECs可形成二维管腔样结构。结论成功建立了人ODMECs的免疫磁珠分离培养方法,并观察到ODMECs具有体外二维管腔样结构。
Objective To establish a magnetic beads sorting system for human ovarian carcinoma-derived microvascular endothelial cells (ODMECs) and observe the formation of two-dimensional tubule-like structure (OMCECs) in vitro. Methods ODMECs were isolated and purified by MACS MicroBeads immunomagnetic bead sorting system combined with CD31 antibody. The obtained ODMECs were identified by morphological, biochemical and phenotypic characteristics and their uptake of acetylated-low-density lipoprotein (DIL-acLDL) and lectin I (UEA-I) Ability to observe the formation of two-dimensional lumen-like structures in an in vitro culture system. Results The purity of ODMECs obtained by flow cytometry was 99.6%, with pebble sign, contact inhibition, expression of CD31, CD105 and FⅧ-Rag; ability of binding to UEA-Ⅰ and uptake of DIL-acLDL; ODMECs were observed during culture Form a two-dimensional lumen-like structure. Conclusion The method of immunomagnetic bead isolation and culture of human ODMECs was successfully established and the OMCECs were observed to have two-dimensional lumen-like structures in vitro.