Yuan Si-Ming, Chen Rong-Liang, Shen Wei-Min, Chen Hai-Ni, Zhou Xiao-Jun
Department of Pathology, Jinling Hospital, Medical School of Nanjing University, Nanjing, Jiangsu, China.
Pediatr Dev Pathol. 2012 Jan-Feb;15(1):5-12. doi: 10.2350/11-01-0959-OA.1. Epub 2011 Jun 20.
Infantile hemangioma grows quickly in the first year of life and regresses slowly to fibrofatty tissue during childhood; mesenchymal stem cells (MSCs) have been reported to contribute to this adipogenesis. Recent studies have shown the perivascular origin of MSCs in multiple organs. We hypothesized that MSCs in hemangioma might also reside in the perivascular region. We isolated MSCs from proliferating hemangioma by their selective adhesion to plastic culture dishes. Mesenchymal stem cells from bone marrow (BM-MSCs) and foreskin-derived fibroblasts were used as controls. Flow cytometry and immunofluorescence staining were used to examine their antigen profiles; in vitro induction of multi-lineage differentiation was performed to test their pluripotency. Platelet-derived growth factor R-β (PDGFR-β), CD133, and peroxisome-proliferator-activated receptor gamma (PPAR-γ) were selected as the markers to observe MSCs in hemangioma by immunohistochemistry staining, with costaining of CD31 and alpha-smooth muscle actin (α-SMA). Hemangioma-derived MSCs (Hem-MSCs) had fibroblast-like morphology; they expressed the MSC markers CD105, CD90, CD29, and vimentin and did not express the hematopoietic/endothelial markers CD45, CD34, CD31, and flt-1; Hem-MSCs also expressed CD133 and PPAR-γ. Most Hem-MSCs expressed PDGFR-β and α-SMA; in contrast, the expression of PDGFR-β and α-SMA in BM-MSCs was very weak. The Hem-MSCs differentiated into adipocytes, osteoblasts, and chondroblasts in vitro. This confirmed their pluripotency. Immunohistochemistry showed the colocalization of PDGFR-β/α-SMA, CD133/α-SMA, and PPAR-γ/α-SMA in the perivascular region. MSCs were successfully obtained from proliferating hemangioma, revealing the perivascular origin of MSCs in hemangioma.
婴儿血管瘤在生命的第一年迅速生长，在儿童期缓慢消退为纤维脂肪组织；据报道，间充质干细胞（MSCs）参与了这种脂肪生成过程。最近的研究表明，多个器官中的MSCs起源于血管周围。我们推测血管瘤中的MSCs可能也存在于血管周围区域。我们通过将增殖期血管瘤中的细胞选择性黏附于塑料培养皿来分离MSCs。将来自骨髓的间充质干细胞（BM-MSCs）和包皮成纤维细胞用作对照。采用流式细胞术和免疫荧光染色检测其抗原谱；进行体外多向分化诱导以测试其多能性。选择血小板衍生生长因子R-β（PDGFR-β）、CD133和过氧化物酶体增殖物激活受体γ（PPAR-γ）作为标志物，通过免疫组织化学染色观察血管瘤中的MSCs，并与CD31和α-平滑肌肌动蛋白（α-SMA）进行共染色。血管瘤来源的MSCs（Hem-MSCs）具有成纤维细胞样形态；它们表达MSCs标志物CD105、CD90、CD29和波形蛋白，不表达造血/内皮标志物CD45、CD34 CD31和flt-1；Hem-MSCs还表达CD133和PPAR-γ。大多数Hem-MSCs表达PDGFR-β和α-SMA；相比之下，BM-MSCs中PDGFR-β和α-SMA的表达非常弱。Hem-MSCs在体外可分化为脂肪细胞、成骨细胞和软骨细胞。这证实了它们的多能性。免疫组织化学显示PDGFR-β/α-SMA、CD133/α-SMA和PPAR-γ/α-SMA在血管周围区域共定位。成功从增殖期血管瘤中获得了MSCs，揭示了血管瘤中MSCs的血管周围起源。
