Graduate School of Life Science, CHA Stem Cell Institute, College of Medicine, CHA University, Gangnam gu, Seoul, Korea.
Cytotherapy. 2011 Feb;13(2):165-78. doi: 10.3109/14653249.2010.512632.
Stem cells have been shown to have a therapeutic effect in several ischemic animal models, including hindlimb ischemia and chronic wound. We examined the wound-healing effect of secretory factors released by human embryonic stem cell (hESC)-derived endothelial precursor cells (EPC) in cutaneous excisional wound models.
hESC-EPC were sorted by CD133/KDR, and endothelial characteristics were confirmed by reverse transcription (RT)-polymerase chain reaction (PCR), Matrigel assay and ac-LDL uptake. Conditioned medium (CM) of hESC-EPC was prepared, and concentrated hESC-EPC CM was applied in a mouse excisional wound model.
hESC-EPC CM accelerated wound healing and increased the tensile strength of wounds after topical treatment and subcutaneous injection. In addition, hESC-EPC CM treatment caused more rapid re-formation of granulation tissue and re-epithelialization of wounds compared with control vehicle medium and CB-EPC CM-treated wounds. In vitro, hESC-EPC CM significantly improved the proliferation and migration of dermal fibroblasts and epidermal keratinocytes. hESC-EPC CM also increased the extracellular matrix synthesis of fibroblasts. Analysis of hESC-EPC CM with a multiplex cytokine array system indicated that hESC-EPC secreted distinctively different cytokines and chemokines, such as epidermal growth factor (EGF), fibroblast growth factor (bFGF), fractalkine, granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-6, IL-8, platelet-derived growth factor-AA (PDGF-AA) and vascular endothelial growth factor (VEGF), which are well known to be important in normal angiogenesis and wound healing.
This study has demonstrated the wound-healing effect of hESC-EPC CM and characterized the spectrum of cytokines released by hESC-EPC that are functionally involved in the wound-healing process. These results suggest that secretory factors released from stem cells could be an important mediator of stem cell therapy in ischemic tissue diseases.
干细胞在多种缺血动物模型中表现出治疗作用，包括后肢缺血和慢性创面。我们研究了人胚胎干细胞（hESC）衍生的内皮前体细胞（EPC）分泌的分泌因子在皮肤切除创面模型中的愈合作用。
通过 CD133/KDR 分选 hESC-EPC，通过逆转录（RT）-聚合酶链反应（PCR）、Matrigel 测定和 ac-LDL 摄取来确认内皮细胞特征。制备 hESC-EPC 条件培养基（CM），并将浓缩的 hESC-EPC CM 应用于小鼠切除创面模型。
hESC-EPC CM 经局部和皮下给药后可加速创面愈合并增加创面的拉伸强度。此外，与对照载体培养基和 CB-EPC CM 处理的创面相比，hESC-EPC CM 处理可使创面更快地形成肉芽组织和再上皮化。体外，hESC-EPC CM 显著促进真皮成纤维细胞和表皮角质形成细胞的增殖和迁移。hESC-EPC CM 还增加了成纤维细胞的细胞外基质合成。通过多功能细胞因子阵列系统分析 hESC-EPC CM 表明，hESC-EPC 分泌独特的细胞因子和趋化因子，如表皮生长因子（EGF）、成纤维细胞生长因子（bFGF）、 fractalkine、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素（IL）-6、IL-8、血小板衍生生长因子-AA（PDGF-AA）和血管内皮生长因子（VEGF），这些因子在正常血管生成和伤口愈合中非常重要。
本研究证实了 hESC-EPC CM 的促愈合作用，并描述了 hESC-EPC 释放的细胞因子谱，这些细胞因子在伤口愈合过程中具有功能作用。这些结果表明，干细胞分泌的分泌因子可能是缺血性组织疾病中干细胞治疗的重要介质。
