Huang Lan, Bischoff Joyce
Vascular Biology Program and Department of Surgery, Boston Children's Hospital, Boston, MA, USA.
Department of Surgery, Harvard Medical School, Boston, MA, USA.
Bio Protoc. 2020 Jan 20;10(2):e3487. doi: 10.21769/BioProtoc.3487.
Infantile hemangioma (IH) is a vascular tumor noted for its excessive blood vessel formation during infancy, glucose-transporter-1 (GLUT1)-positive staining of the blood vessels, and its slow spontaneous involution over several years in early childhood. For most children, IH poses no serious threat because it will eventually involute, but a subset can destroy facial structures and impair vision, breathing and feeding. To unravel the molecular mechanism(s) driving IH-specific vascular overgrowth, which to date remains elusive, investigators have studied IH histopathology, the cellular constituents and mRNA expression. Hemangioma endothelial cells (HemEC) were first isolated from surgically removed IH specimens in 1982 by Mulliken and colleagues ( Mulliken , 1982 ). Hemangioma stem cells (HemSC) were isolated in 2008, hemangioma pericytes in 2013 and GLUT1-positive HemEC in 2015. Indeed, as we describe here, it is possible to isolate HemSC, GLUT1-positive HemEC, GLUT1-negative HemEC and HemPericytes from a single proliferating IH tissue specimen. This is accomplished by sequential selection using antibodies against specific cell surface markers: anti-CD133 to select HemSC, anti-GLUT1 and anti-CD31 to select HemECs and anti-PDGFRβ to select HemPericytes. IH-derived cells proliferate well in culture and can be used for and vasculogenesis and angiogenesis assays.
婴儿血管瘤（IH）是一种血管肿瘤，其特点是在婴儿期血管过度形成、血管葡萄糖转运蛋白1（GLUT1）呈阳性染色，并且在儿童早期会在数年时间里缓慢自然消退。对大多数儿童来说，婴儿血管瘤不会构成严重威胁，因为它最终会消退，但有一部分婴儿血管瘤会破坏面部结构并损害视力、呼吸和进食功能。为了阐明导致婴儿血管瘤特异性血管过度生长的分子机制（迄今为止仍不清楚），研究人员对婴儿血管瘤的组织病理学、细胞成分和mRNA表达进行了研究。1982年，Mulliken及其同事首次从手术切除的婴儿血管瘤标本中分离出血管瘤内皮细胞（HemEC）（Mulliken，1982）。2008年分离出血管瘤干细胞（HemSC），2013年分离出血管瘤周细胞，2015年分离出GLUT1阳性的HemEC。事实上，正如我们在此所描述的，从单个增殖性婴儿血管瘤组织标本中分离出HemSC、GLUT1阳性的HemEC、GLUT1阴性的HemEC和Hem周细胞是可能的。这是通过使用针对特定细胞表面标志物的抗体进行顺序选择来实现的：抗CD133用于选择HemSC，抗GLUT1和抗CD31用于选择HemEC，抗PDGFRβ用于选择Hem周细胞。源自婴儿血管瘤的细胞在培养中增殖良好，可用于血管生成和血管新生测定。
