内皮祖细胞在裸鼠体内再现人血管瘤的生物学特性研究
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摘要
目的:研究人血管瘤内皮祖细胞(EPcs)在裸鼠体内分化情况,进一步揭示其生物学特性。方法:组织块法体外培养增生期血管瘤内皮祖细胞(经CD31、CD34及Ⅷ因子相关抗原鉴定),浓集后分别种植于裸鼠双侧胸壁、腰部皮下,动态观察移植瘤生长情况及组织学变化,并用血管内皮细胞特异性标志物鼠抗人CD31、CD34免疫组化单克隆抗体染色研究所成瘤体的同源性。结果:体外培养血管瘤内皮祖细胞生长良好,细胞计数为3×10~8个/培养瓶,CD31、CD34及Ⅷ因子相关抗原免疫组化检测阳性表达,裸鼠皮下植入后所形成的瘤体与原血管瘤生物学特点及病理特征相似,经CD31、CD34鉴定瘤体来源于人血管瘤内皮细胞。结论:内皮祖细胞是血管瘤发生的细胞来源,此可作为血管瘤的动物模型,为血管瘤的研究提供较理想平台。
Objective:To study the differentiation of human hemangioma endothelial progenitor cells(EPcs) in nude mice,and further reveal its biological characteristics.Methods:The hemangioma endothelial progenitor cells in proliferative stage were cultured in vitro,identified by CD31,CD34 and Ⅷ-factor related antigens,and subcutaneously inoculated into the bilateral chest wall and waist of nude mice.The growth characteristics and histologic changes of transplanted tumors were regularly observed.The transplanted tumor′s homology was identified by immunohistochemical staining of the vascular endothelial cell specific marker CD31 and CD34.Results:The growth of EPcs was good in vitro,the number of EPcs was 3×10~8,and the positive expressions of CD31,CD34 and Ⅷ-factor related antigen were detected by immunohistochemistry.The biological and pathological characteristics between tumor formation after subcutaneous implantation and original hemangioma were similar in nude mice.The tumor identified by CD31 and CD34 was derived from human hemangioma endothelial cells.Conclusions:The cell source of hemangioma is derived from EPCs,which can be used as the animal models of hemangioma,and provide an ideal platform for the study of hemangioma.
Objective:To study the differentiation of human hemangioma endothelial progenitor cells(EPcs) in nude mice,and further reveal its biological characteristics.Methods:The hemangioma endothelial progenitor cells in proliferative stage were cultured in vitro,identified by CD31,CD34 and Ⅷ-factor related antigens,and subcutaneously inoculated into the bilateral chest wall and waist of nude mice.The growth characteristics and histologic changes of transplanted tumors were regularly observed.The transplanted tumor′s homology was identified by immunohistochemical staining of the vascular endothelial cell specific marker CD31 and CD34.Results:The growth of EPcs was good in vitro,the number of EPcs was 3×10~8,and the positive expressions of CD31,CD34 and Ⅷ-factor related antigen were detected by immunohistochemistry.The biological and pathological characteristics between tumor formation after subcutaneous implantation and original hemangioma were similar in nude mice.The tumor identified by CD31 and CD34 was derived from human hemangioma endothelial cells.Conclusions:The cell source of hemangioma is derived from EPCs,which can be used as the animal models of hemangioma,and provide an ideal platform for the study of hemangioma.
引文
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[2] TAN ST,HASANQ,VELICKOVIC M,et al.A Novel in vitro human model of hemangioma[J].Mod Pathol,2000,13(1):92.
[3] HOAK JC,WARNER ED,CHENG HF,et al.Hemangioma with thrombocytopenia and microangiopathic anemia (Kasabach-Merritt syndrome):an animal model[J].J Lab Clin Med,1971,77(6):941.
[4] PRIMO L,ROCA C,FERRANDI C,et al.Human endothelial cells expressing polyma middle T induce tumors [J].Oncogene,2000,19(32):3632.
[5] TANG Y,LIU W,YU S,et al.A novel in vivo model of human hemangioma:xenograft of human hemangioma tissue on nude mice[J].Plast Reconstr Surg,2007,120(4):869.
[6] 虎小毅,杨壮群,宋勇,等.细胞悬液接种法血管瘤裸鼠动物模型的建立[J].中国美容医学,2008,17(7):1012.
[7] MEININGER CJ,BRIGHTMAN SE,KELLY K A,et al.Increased stem cell factor release by hemangioma-derived endothelial cells[J].Lab Invest,1995,72(2):166.
[8] PETERSON AW,CALDWELL DJ,RIOJA AY,et al.Vasculogenesis and angiogenesis in modular collagen-fibrin microtissues[J].Biomater Sci,2014,2(10):1497.
[9] 稂翠玲,孙斌,雷红召,等.血管瘤血管内皮细胞体外分离纯化方法的优化[J].中国美容医学,2018,27(4):65.
[10] 李攀登,陈润芳,张莉,等.婴幼儿增生期血管瘤内皮细胞观察:培养、鉴定及生长状况[J].中国组织工程研究,2015,19(51):8276.
[11] CHALAJOUR F,TREEDE H,EBRAH IMNEJAD A,et al.Angiogenic activation of vascular endothelial cells in aortic valve stenosis[J].Exp Cell Res,2004,298(2):455.
[12] BALAN S,KALE VP,LIMAYE LS.A simple two-step culture system for the large-scale generation of mature and functional dendritic cells from umbilical cord blood CD34+ cells[J].Transfusion,2009,49(10):13.
[13] 米萱.自噬抑制2-DG诱导的内皮细胞凋亡及自噬在婴幼儿血管瘤中的活性研究[D].西安:第四军医大学,2016.
[14] HARBI S,PARK H,GREGORY M,et al.Arrested development:infantile hemangioma and the stem cell teratogenic hypothesis[J].Lymphat Res Biol,2017,15(2):153.
[15] 徐艳朋,俞松,彭岗,等.Raf-1和p-ERK在血管瘤裸鼠移植模型中的表达及意义[J].中华小儿外科杂志,2015,36(12):936.