Conditioned medium from umbilical cord mesenchymal stem cells induces migration and angiogenesis.
Conditioned medium from umbilical cord mesenchymal stem cells induces migration and angiogenesis.
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脐带间充质干细胞的条件培养基诱导迁移和血管生成
DOI：
10.3892/mmr.2015.3409
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发表时间：
2015-07
影响因子：
3.4
通讯作者：
Li H
中科院分区：
文献类型：
--
作者：
Shen C;Lie P;Miao T;Yu M;Lu Q;Feng T;Li J;Zu T;Liu X;Li H
脐带间充质干细胞(UC-MSCs)已被认为可用于各种临床应用，但主要的局限性包括缺乏器官特异性蓄积和移植细胞存活率低。在本研究中，假设UC-MSCs的旁分泌作用可能通过促进干/祖细胞的特异性归巢和将其驱动到受损区域的整体能力来促进基于干细胞的组织修复和再生。采用液体芯片和ELISA法分析UC-MSCs来源的条件培养液(UC-CM)。体外培养人脐静脉内皮细胞(HUVECs)和UC-MSCs，观察UC-CM的血管生成特性。随后，用PKH26标记UC-MSCs、HUVECs和成纤维细胞进行体内细胞迁移实验。采用逆转录-定量聚合酶链式反应和流式细胞术检测UC-MSCs、HUVECs和成纤维细胞中C-X-C趋化因子受体4(CXCR4)、C-C趋化因子受体2(CCR2)和c-met的表达水平。体外培养UC-CM，观察基质细胞衍生因子-1(SDF-1)、单核细胞趋化蛋白-1(MCP-1)和肝细胞生长因子(HGF)对细胞迁移的影响。结果表明，UC-MSCs除分泌血管生成因子、血管细胞黏附蛋白-1、白介素8、胰岛素样生长因子和血管内皮生长因子外，还分泌不同的细胞因子和趋化因子，包括SDF-1、MCP-1和HGF。UC-CM培养的UC-MSCs和HUVECs较Dulbecco改良的Eagle‘s培养液培养的UC-MSCs和HUVECs的总管长明显增加。体内细胞迁移实验表明，UC-CM对UC-MSCs和HUVECs具有趋化作用。体外Matrigel迁移和划痕愈合实验表明，UC-CM以剂量依赖的方式增加CXCR4阳性和/和CCR2阳性细胞的迁移。此外，在基于抗体的封闭迁移条件下筛选了不同的分子。结果表明，SDF-1/CXCR4和MCP-1/CCR2轴参与了UC-CM的趋化活性，提示UC-CM的有效旁分泌因子是一个大的复合体，而不是一个单一的因素。本研究结果支持UC-MSCs释放可溶性因子的假说，这可能扩大干细胞的治疗适用性。
Umbilical cord mesenchymal stem cells (UC-MSCs) have been suggested as a candidate for various clinical applications, however, major limitations include the lack of organ-specific accumulation and low survival rates of transplanted cells. In the present study, it was hypothesized that the paracrine effects of UC-MSCs may enhance stem cell-based tissue repair and regeneration by promoting the specific homing of stem/progenitor cells and the overall ability to drive them to the damaged area. UC-MSCs-derived conditioned medium (UC-CM) was analyzed using liquid chip and ELISA techniques. In vitro tube formation assays of human umbilical vein endothelial cells (HUVECs) and UC-MSCs were then performed to assess the angiogenic properties of UC-CM. Subsequently, UC-MSCs, HUVECs and fibroblasts were labeled with PKH26 for an in vivo cell migration assay. The expression levels of C-X-C chemokine receptor 4 (CXCR4), C-C chemokine receptor 2 (CCR2) and c-met were determined in the UC-MSCs, HUVECs and fibroblasts using reverse transcription-quantitative polymerase chain reaction and flow cytometry. UC-CM was incubated with or without antibodies, and the contribution of stromal cell-derived factor 1 (SDF-1), monocyte chemotactic protein 1 (MCP-1) and hepatocyte growth factor (HGF) on the migration of cells was investigated in vitro. The results demonstrated that UC-MSCs secreted different cytokines and chemokines, including increased quantities of SDF-1, MCP-1 and HGF, in addition to the angiogenic factors, vascular cell adhesion protein-1, interleukin-8, insulin-like growth factor-1 and vascular endothelial growth factor. The total lengths of the tubes were significantly increased in the UC-MSCs and HUVECs incubated in UC-CM compared with those incubated in Dulbecco’s modified Eagle’s medium. In vivo cell migration assays demonstrated that UC-CM was a chemotactic stimulus for the UC-MSCs and HUVECs. In vitro Matrigel migration and scratch healing assays demonstrated that UC-CM increased the migration of CXCR4-postive or/and CCR2-positive cells in a dose-dependent manner. In addition, different molecules were screened under antibody-based blocking migration conditions. The data revealed that the SDF-1/CXCR4 and MCP-1/CCR2 axes were involved in the chemoattractive activity of UC-CM and suggested that the effective paracrine factor of UC-CM is a large complex rather than a single factor. The results of the present study supported the hypothesis that UC-MSCs release soluble factors, which may extend the therapeutic applicability of stem cells.
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影响因子：
3.3
作者：
Cavallari, Giuseppe;Olivi, Elena;Ventura, Carlo
通讯作者：
Ventura, Carlo
DOI：
10.1152/ajpheart.00901.2003
发表时间：
2004-03-01
影响因子：
4.8
作者：
Ashrafpour, H;Huang, N;Pang, CY
通讯作者：
Pang, CY
影响因子：
9.5
作者：
Ma, J;Ge, JB;Zou, YZ
通讯作者：
Zou, YZ
影响因子：
20.3
作者：
Peled, A;Kollet, O;Lapidot, T
通讯作者：
Lapidot, T
DOI：
10.1097/01.asn.0000089563.63641.a8
发表时间：
2003-10-01
影响因子：
13.6
作者：
Furuichi, K;Wada, T;Yokoyama, H
通讯作者：
Yokoyama, H