Lyu Dongze, Ying Hanru, Chang Lei, Ma Gang, Lin Xiaoxi
Zhonghua Zheng Xing Wai Ke Za Zhi. 2016 Jul;32(4):293-8.
To establish a reliable method of isolation and culture of infantile hemangioma stem cells (HemSCs).
Proliferating infantile hemangioma specimens were digested with collagenase to form a single cell suspension. The HemSCs were isolated with anti-CD133 MicroBeads, and were incubated in fibronectin coated 96-well plates with EBM-2 (10％ FBS). HemSCs were identified by morphological characteristics, flow cytometry, cell tubule formation assay, osteoinductive and adipogenic differentiation assay, and subcutaneous tumor formation assay.
This method enables the rapid isolation of HemSCs which demonstrated typical mesenchymal stem cell morphology in culture.CD133 (+) HemSCs expressed CD29 (99.5％),CD44 (97.9％),CD90 (87.6％) and CD105 (98.5％),but barely expressed CD31 (0.2％),CD34 (0.1％),CD45 (0.1％) and CD144 (0.1％).These cells could differentiate into osteoblasts and adipocytes,and could form vascular wall like structure in vitro. When implanted into subcutaneous of the nude mice, the cells can develop into hemangioma like lesion histologically.
This technique can effectively isolate HemSCs from the proliferative hemangioma. These cells could be further used to reveal the charaeteristics of HemSCs, as well as for further study of widespread application.
建立一种可靠的婴幼儿血管瘤干细胞（HemSCs）分离培养方法。
将增殖期婴幼儿血管瘤标本用胶原酶消化形成单细胞悬液。用抗CD133微珠分离HemSCs，并接种于纤连蛋白包被的96孔板中，加入含10％胎牛血清的EBM-2培养基培养。通过形态学特征、流式细胞术、细胞小管形成实验、成骨和成脂分化实验以及皮下肿瘤形成实验对HemSCs进行鉴定。
该方法能够快速分离出HemSCs，其在培养中表现出典型的间充质干细胞形态。CD133（+）HemSCs表达CD29（99.5％）、CD44（97.9％）、CD90（87.6％）和CD105（98.5％），但几乎不表达CD31（0.2％）、CD34（0.1％）、CD45（0.1％）和CD144（0.1％）。这些细胞可分化为成骨细胞和成脂细胞，并能在体外形成血管壁样结构。将其植入裸鼠皮下，组织学检查显示可发展为血管瘤样病变。
该技术可有效从增殖期血管瘤中分离出HemSCs。这些细胞可进一步用于揭示HemSCs的特性，以及用于更广泛应用的深入研究。
