Stable expression of angiopoietin-1 and other markers by cultured pericytes: Phenotypic similarities to a subpopulation of cells in maturing vessels during later stages of angiogenesis in vivo
Stable expression of angiopoietin-1 and other markers by cultured pericytes: Phenotypic similarities to a subpopulation of cells in maturing vessels during later stages of angiogenesis in vivo
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DOI：
10.1038/labinvest.3780433
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发表时间：
2002-04-01
影响因子：
5
通讯作者：
Dvorak, HF
中科院分区：
文献类型：
--
作者：
Sundberg, C;Kowanetz, M;Dvorak, HF
周细胞一直是难以研究的细胞，因为它们在体外不能保持其特征表型，并且仅在培养几天后就开始表达成纤维细胞标记物。我们现在报道了使用免疫亲和磁珠方法与周细胞表面神经节苷脂特异性的 3G5 IgM 单克隆抗体从混合细胞群中分离、纯化、培养和再纯化人真皮周细胞的方法。这些纯化的细胞可以在培养物中扩增，并且它们的周细胞表型可维持长达 8 天。此外，它们强烈表达血管生成素-1 (Ang-1)，但不表达血管生成素-2、Tie-1 或 Tie-2；相反，真皮微血管内皮细胞表现出相反的表达模式。这些发现很重要，因为内皮细胞和周细胞非常接近，通常很难确定原位表达这些蛋白质的特定细胞类型。将我们的体外研究结果扩展到体内血管生成的两种模型，我们证明了在皮肤伤口愈合和血管通透因子/血管内皮生长因子诱导的血管生成的后期阶段，成熟的微血管中出现了表达Ang-1的细胞亚群。我们的结果提供了强有力的证据，表明 Ang-1 在体外和体内由周细胞表达，并且 Ang-1 在发育中血管成熟中的作用可以扩展到成体组织中血管生成后的血管成熟。
Pericytes have been difficult cells to study because they do not maintain their characteristic phenotype in vitro, and they begin to express fibroblast markers after only a few days in culture. We now report methods for the isolation, purification, culture, and repurification of human dermal pericytes from mixed cell populations using an immunoaffinity-magnetic bead approach coupled with the 3G5 IgM monoclonal antibody that is specific for a pericyte surface ganglioside. These purified cells could be expanded in culture, and they maintained their pericyte phenotype for up to 8 days. In addition, they strongly expressed angiopoietin-1 (Ang-1) but not angiopoietin-2, Tie-1, or Tie-2; in contrast, dermal microvascular endothelial cells exhibited a reciprocal expression pattern. These findings are important because the close proximity of endothelial cells and pericytes has often made it difficult to determine with certainty the specific cell type(s) that expressed each of these proteins in situ. Extending our in vitro findings to two models of angiogenesis in vivo, we demonstrated a subpopulation of Ang-1-expressing cells that appeared in maturing microvessels during later stages of cutaneous wound healing and vascular permeability factor/vascular endothelial growth factor-induced angiogenesis. Our results provide strong evidence that Ang-1 is expressed by pericytes in vitro and in vivo and that the role proposed for Ang-1 in vessel maturation in development can be extended to vessel maturation after angiogenesis in adult tissues.