婴幼儿血管瘤细胞系XPTS-1和动物模型的建立

目的 建立婴幼儿血管瘤源性血管内皮细胞系(hemangioma-derived endothelial cell line,HemEC)及其动物模型.方法 采用组织块法进行HemEC体外培养,制作HemEC生长曲线,免疫组化法行Ⅷ因子相关抗原鉴定,流式细胞仪鉴定HemEC的血管内皮生长因子受体2(vascular endothelial growth factor receptor2,VEGFR-2)的表达,分析HemEC染色体核型,采用异体移植的方法将HemEC接种于裸鼠皮下,观察肿瘤的生长情况,进行组织病理学检查.结果 培养的细胞生长活跃,传到第16代时细胞增殖增快,自发永生化,共传代培养70代以上,体外培养时间1年,冻存、复苏多次,细胞仍生长良好,具有HemEC的形态学特征,细胞群体倍增时间约为22 h,Ⅷ因子和VEGFR-2均阳性表达,HemEC的染色体表现为非正常的二倍体特征,染色体数介于二倍体和三倍体之间,将其命名为XPTS-1.该婴幼儿血管瘤动物模型的组织病理学特征与婴幼儿血管瘤的组织病理学特征基本一致.结论 本研究建立的HemEC自发转化、永生化,其形态学特征和生物学特性符合HemEC的特性,既具有一般转化细胞系的饱和密度生长、接触性抑制、停泊依赖性生长等特性,又具有致瘤性等特性,符合婴幼儿血管瘤组织病理学特性,该模型的组织病理学特征与婴幼儿血管瘤的组织病理学特征基本一致.

Abstract：

Objective To establish an immortalized human infancy hemangioma-derived endothelial cell line (HemEC) and animal model of human infancy hemangioma. Methods Hemangioma-derived endothelial cells from specimen of human infancy hemangioma were cultured in vitro and monocloed, and then its growth curve was made, karyomorphism of chromosome analyzed, morphologic characteristics observe,factor Ⅷ related antigen identified by immunohistochemical method. Vascular endothelial growth factor receptor 2(VEGFR-2) was detected by flow cytometry. HemEC were inoculated subcutaneously in athymicmouse to establish animal model of infancy hemangioma. The animal model was observed closely and its pathological characteristic was also studied. Results The cultural cells grew active, and immortalized spontaneously when they were subcultured on sixteenth generation. This cell line was cultivated for more than 70 times within one year and in good condition after freezing and resuscitating once and again, and had the morphologic character of HemEC. The cell population doubling time was 22 h. Factor Ⅷ and VEGFR-2 were expressed positively. Karyo type analysis of the cell line showed abnormal diploid with the modal chromosomal number varying between diploid and triploid. The cell line was then named XPTS-1. The animal model of infancy hemangioma was successfully established and its character of histopathology was similar with that of infancy hemangioma. Conclusions The cell line of HemEC was successfully established and immortalized spontaneously, and had the morphologic and biological character of HemEC. The animal model of infancy hemangioma was successfully established and showed the character of histopathology similar with that of infancy hemangioma.

Establishment of human infancy hemangioma-derived endothelial cell line XPTS-1 and animal model of human infancy hemangioma

Objective To establish an immortalized human infancy hemangioma-derived endothelial cell line (HemEC) and animal model of human infancy hemangioma. Methods Hemangioma-derived endothelial cells from specimen of human infancy hemangioma were cultured in vitro and monocloed, and then its growth curve was made, karyomorphism of chromosome analyzed, morphologic characteristics observe,factor Ⅷ related antigen identified by immunohistochemical method. Vascular endothelial growth factor receptor 2(VEGFR-2) was detected by flow cytometry. HemEC were inoculated subcutaneously in athymicmouse to establish animal model of infancy hemangioma. The animal model was observed closely and its pathological characteristic was also studied. Results The cultural cells grew active, and immortalized spontaneously when they were subcultured on sixteenth generation. This cell line was cultivated for more than 70 times within one year and in good condition after freezing and resuscitating once and again, and had the morphologic character of HemEC. The cell population doubling time was 22 h. Factor Ⅷ and VEGFR-2 were expressed positively. Karyo type analysis of the cell line showed abnormal diploid with the modal chromosomal number varying between diploid and triploid. The cell line was then named XPTS-1. The animal model of infancy hemangioma was successfully established and its character of histopathology was similar with that of infancy hemangioma. Conclusions The cell line of HemEC was successfully established and immortalized spontaneously, and had the morphologic and biological character of HemEC. The animal model of infancy hemangioma was successfully established and showed the character of histopathology similar with that of infancy hemangioma.