Podoplanin-expressing cells derived from bone marrow play a crucial role in postnatal lymphatic neovascularization.
Podoplanin-expressing cells derived from bone marrow play a crucial role in postnatal lymphatic neovascularization.
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DOI：
10.1161/circulationaha.110.941468
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发表时间：
2010-10-05
期刊：
影响因子：
37.8
通讯作者：
Yoon YS
中科院分区：
文献类型：
--
作者：
Lee JY;Park C;Cho YP;Lee E;Kim H;Kim P;Yun SH;Yoon YS
新出现的证据表明骨髓（BM）细胞对淋巴管形成的贡献；然而，具有淋巴内皮祖细胞（LEPC）功能的细胞的确切表型尚未确定。在这里，我们研究脑卒中来源的lepc的身份及其在淋巴新生血管中的作用。在VEGFA、VEGFC和EGF存在下培养bm -单核细胞（MNCs），导致淋巴内皮细胞（LEC）标志物的表达。其中podoplanin+细胞采用磁标记细胞分离系统（MACS）分离，FACS和免疫细胞化学鉴定。这些podoplanin+细胞高度表达LECs、造血谱系和干细胞/祖细胞的标记物，并在进一步培养后产生LECs。我们进一步证实，podoplanin+细胞在正常小鼠的骨髓和外周血（PB）中少量存在，但在肿瘤植入等淋巴管生成刺激下显著增加（15倍）。接下来，为了评估podoplanin+细胞在体内形成新淋巴管的潜力，我们将培养分离或新分离的bm来源的podoplanin+细胞注射到伤口和肿瘤模型中。免疫组化结果显示，注射细胞被纳入淋巴管系统，呈现LEC表型，组织中淋巴管密度增加，提示淋巴管发生。在共培养过程中，Podoplanin+细胞也表达了高水平的淋巴管生成细胞因子，并增加了LECs的增殖，表明其具有淋巴管生成或旁分泌的作用。我们的研究结果提供了令人信服的证据，证明bm来源的podoplanin+细胞（一种以前未被识别的细胞类型）具有lepc的功能，并通过淋巴管生成和淋巴管生成参与出生后淋巴管新生。
Emerging evidence has suggested a contribution of bone marrow (BM) cells to lymphatic vessel formation; however, the exact phenotype of the cells with lymphatic endothelial progenitor cell (LEPC) function has yet to be identified. Here we investigate the identity of BM-derived LEPCs and their role in lymphatic neovascularization. Culture of BM-mononuclear cells (MNCs) in the presence of VEGFA, VEGFC and EGF resulted in expression of lymphatic endothelial cell (LEC) markers. Among these cells, podoplanin+ cells were isolated by magnetic-labeled cell separation system (MACS) and characterized by FACS and immunocytochemistry. These podoplanin+ cells highly express markers for LECs, hematopoietic lineages, and stem/progenitor cells, and upon further cultivation, generate LECs. We further confirmed that podoplanin+ cells exist in small numbers in BM and peripheral blood (PB) of normal mice, but are significantly (15 fold) augmented upon lymphangiogenic stimuli such as tumor implantation. Next, to evaluate the potential of podoplanin+ cells for the formation of new lymphatic vessels in vivo, we injected culture-isolated or freshly isolated BM-derived podoplanin+ cells into wound and tumor models. Immunohistochemistry demonstrated that the injected cells were incorporated into the lymphatic vasculature, displayed LEC phenotypes, and increased lymphatic vascular density in tissues, suggesting lymphvasculogenesis. Podoplanin+ cells also expressed high levels of lymphangiogenic cytokines and increased proliferation of LECs during co-culture, suggesting a lymphangiogenic or paracrine role. Our results provide compelling evidence that BM-derived podoplanin+ cells, a previously unrecognized cell type, function as LEPCs and participate in postnatal lymphatic neovascularization through both lymphvasculogenesis and lymphangiogenesis.