Takeda Kohsuke, Sowa Yoshihiro, Nishino Kenichi, Itoh Kyoko, Fushiki Shinji
From the Departments of *Surgery, Division of Plastic and Reconstructive Surgery, and †Pathology and Applied Neurobiology, Graduate School of Medical Sciences, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Ann Plast Surg. 2015 Jun;74(6):728-36. doi: 10.1097/SAP.0000000000000084.
Adipose-derived stem cells (ADSCs) are a promising new therapeutic modality for several diseases and have been applied to various clinical fields because of their multidifferentiation potential and capacity for growth-factor secretion. Recently, 2 in vivo studies showed ADSCs to have potential applications in lymphedema therapy. However, it remains unclear whether ADSCs have direct effects on lymphatic endothelial cells (LECs). In this study, human LECs were treated with murine ADSC-derived conditioned media. Changes in LEC proliferation, migration, and tube formation were assessed by WST-8 assay, transwell chamber assay, and Matrigel-based tube formation assay, respectively, with recombinant human vascular endothelial growth factor-C used as a positive control. Additionally, the expression of several lymphangiogenic factors in ADSCs was examined by quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Factors secreted by ADSCs induced LEC proliferation, migration, and tube formation more potently than recombinant human vascular endothelial growth factor-C. We confirmed by quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay that some of the lymphangiogenic factors of ADSCs were dramatically up-regulated under serum-starved conditions. These data indicate that ADSCs could directly contribute to lymphangiogenesis via secretory factors in vitro and may thus provide a therapeutic modality for patients with lymphedema.
脂肪来源干细胞（ADSCs）是一种对多种疾病颇具前景的新型治疗方式，因其具有多向分化潜能和分泌生长因子的能力，已被应用于各个临床领域。最近，两项体内研究表明ADSCs在淋巴水肿治疗中具有潜在应用价值。然而，ADSCs是否对淋巴管内皮细胞（LECs）有直接作用仍不清楚。在本研究中，用人鼠ADSC来源的条件培养基处理人LECs。分别通过WST-8测定、Transwell小室测定和基于基质胶的管形成测定评估LEC增殖、迁移和管形成的变化，以重组人血管内皮生长因子-C作为阳性对照。此外，通过定量逆转录-聚合酶链反应和酶联免疫吸附测定检测ADSCs中几种淋巴管生成因子的表达。ADSCs分泌的因子比重组人血管内皮生长因子-C更有效地诱导LEC增殖、迁移和管形成。我们通过定量逆转录-聚合酶链反应和酶联免疫吸附测定证实，在血清饥饿条件下，ADSCs的一些淋巴管生成因子显著上调。这些数据表明，ADSCs在体外可通过分泌因子直接促进淋巴管生成，因此可能为淋巴水肿患者提供一种治疗方式。
