Department of Pediatrics, Indiana University School of Medicine, Indianapolis, Indiana, USA.
Invest Ophthalmol Vis Sci. 2010 Aug;51(8):3943-9. doi: 10.1167/iovs.09-4970. Epub 2010 Mar 17.
To test the hypothesis that the robust expansion of bovine corneal endothelial cells (BCECs) in vitro is due to the presence of individual endothelial cells with various levels of proliferative potential.
BCECs and bovine vascular endothelial cells (ECs) derived from aorta, coronary artery, and pulmonary artery were cultivated in optimized medium. These cell populations were confirmed by morphologic features, functional assays, and gene expression profiles. Moreover, ECs were plated in a single-cell clonogenic assay to evaluate colony-forming ability.
Both corneal and vascular ECs were confirmed to be pure populations of endothelium uncontaminated with hematopoietic cells. A complete hierarchy of endothelial colony-forming cells (ECFCs) was identified in BCECs by a single-cell clonogenic assay. The distribution of the various types of ECFCs was similar to the control ECs removed from the systemic vessels.
Cultured BCECs display clonal proliferative properties similar to those of vascular ECs.
验证假设，即牛角膜内皮细胞（BCECs）在体外的大量扩增是由于存在具有不同增殖潜能的单个内皮细胞。
在优化的培养基中培养 BCECs 和源自主动脉、冠状动脉和肺动脉的牛血管内皮细胞（ECs）。通过形态特征、功能测定和基因表达谱来确认这些细胞群体。此外，将 ECs 铺板于单细胞克隆形成测定中以评估集落形成能力。
角膜和血管 ECs 均被证实为纯净的内皮细胞群体，未被造血细胞污染。通过单细胞克隆形成测定，在 BCECs 中鉴定出完整的内皮细胞集落形成细胞（ECFCs）层次。各种类型的 ECFCs 的分布与从系统性血管中去除的对照 ECs 相似。
培养的 BCECs 显示出与血管 ECs 相似的克隆增殖特性。
