Human Breast Adipose-Derived Stem Cells Transfected with the Stromal Cell-Derived Factor-1 Receptor CXCR4 Exhibit Enhanced Viability in Human Autologous Free Fat Grafts
Human Breast Adipose-Derived Stem Cells Transfected with the Stromal Cell-Derived Factor-1 Receptor CXCR4 Exhibit Enhanced Viability in Human Autologous Free Fat Grafts
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DOI：
10.1159/000366404
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发表时间：
2014-11
影响因子：
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通讯作者：
Fang-tian Xu;Hong-Mian Li;Q. Yin;Da-lie Liu;Hua Nan;Pei-ran Zhao;Shuang-wu Liang
中科院分区：
文献类型：
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作者：
Fang-tian Xu;Hong-Mian Li;Q. Yin;Da-lie Liu;Hua Nan;Pei-ran Zhao;Shuang-wu Liang
背景资料：自体游离脂肪组织移植的主要并发症是脂肪缺血性坏死引起的脂肪吸收和钙化。自体游离脂肪移植后促进移植物新生血管形成可能会减少这些结果。在成年期，基质细胞衍生因子-1（SDF-1）及其膜受体C-X-C趋化因子受体4（CXCR 4）参与多种干细胞类型的归巢和迁移、新血管形成和细胞增殖。我们假设CXCR 4可能通过募集内皮祖细胞（EPCs）来改善游离脂肪组织移植物的长期存活，因此可能改善移植物的血管重建。在这项研究中，我们的目的是确定与CXCR 4基因转染的人乳腺脂肪源性干细胞（HBASCs）对人自体游离脂肪移植在裸鼠中的存活率的影响。研究方法：将人乳腺脂肪来源的干细胞（HBASC）离体扩增3代，用绿色荧光蛋白（GFP）标记并用CXCR 4转染或保持未转染。将自体脂肪组织与GFP标记的、CXCR 4转染的HBASCs（A组）、GFP标记的HBASCs（B组）、已知的血管生成促进剂VEGF（C组）或培养基（D组）混合，然后以随机方式在4个点皮下注射到32只裸鼠中。6个月后，评估移植组织体积和组织学，并通过计数毛细血管来量化新生血管形成。采用实时定量PCR（qPCR）分析移植物中CXCR 4和SDF-1α mRNA的表达。结果：对照组（D组）移植物体积存活率为28.3 ± 4.5%。混合CXCR 4转染的（A组）和未转染的（B组）HBASC显著增加了移植体积存活率（分别为79.5 ± 8.3%和67.2 ± 5.9%），而VEGF转染的HBASC（C组）效果较差（41.2 ± 5.1%）。组织学分析显示，与对照移植物不同，两种类型的HBASCs处理的移植物主要由脂肪组织组成，并且还呈现出显著较少的脂肪坏死和纤维化。CXCR 4转染的HBASC处理的移植物具有比其他移植物显著更高的毛细血管密度，并显示GFP和CD 31双阳性细胞（即，ASC衍生的内皮细胞）。转染CXCR 4的HBASCs移植物中CXCR 4和SDF-1α的mRNA表达明显高于其他3种移植物。结论：我们的数据表明，HBASCs可以提高移植的游离脂肪组织的存活率和质量。此外，CXCR 4转染这些HBASCs可以增强这种效果。由于内皮祖细胞（EPCs）的募集和移植物再血管化的增加，血管生成的刺激和脂肪细胞凋亡的减少是CXCR 4转染的HBASCs治疗后游离脂肪移植物长期存活改善的潜在机制。
Background: The main complication of autologous free fat tissue transplantation is fat resorption and calcification due to the ischemic necrosis of fat. The promotion of transplant neovascularization soon after autologous free fat grafts may reduce these outcomes. In adulthood, stromal cell-derived factor-1 (SDF-1) and its membrane receptor C-X-C chemokine receptor type 4 (CXCR4) are involved in the homing and migration of multiple stem cell types, neovascularization, and cell proliferation. We hypothesized that CXCR4 may improve the long-term survival of free fat tissue transplants by recruiting endothelial progenitor cells (EPCs) and may therefore improve graft revascularization. In this study, we aimed to determine the effect of human breast adipose-derived stem cells (HBASCs) transfected with the CXCR4 gene on the survival rate of human autologous free fat transplants in nude mice. Methods: Human breast adipose-derived stem cells (HBASCs) were expanded ex vivo for 3 passages, labeled with green fluorescent protein (GFP) and transfected with CXCR4 or left untransfected. Autologous fat tissues were mixed with the GFP-labeled, CXCR4-transfected HBASCs (group A), GFP-labeled HBASCs (group B), the known vascularization-promoting agent VEGF (group C), or medium (group D) and then injected subcutaneously into 32 nude mice at 4 spots in a random fashion. Six months later, the transplanted tissue volume and histology were evaluated, and neo-vascularization was quantified by counting the capillaries. CXCR4 and SDF-1α mRNA expression in the transplants was determined using real-time quantitative PCR analysis (qPCR). Results: The data revealed that the control (group D) transplant volume survival was 28.3 ± 4.5%. Mixing CXCR4-transfected (group A) and untransfected (group B) HBASCs significantly increased transplant volume survival (79.5 ± 8.3% and 67.2 ± 5.9%, respectively), whereas VEGF-transfected HBASCs (group C) were less effective (41.2 ± 5.1%). Histological analysis revealed that both types of HBASCs-treated transplants consisted predominantly of adipose tissue, unlike the control transplants, and also presented significantly less fat necrosis and fibrosis. The CXCR4-transfected HBASCs-treated transplants had a significantly higher capillary density than did the other transplants and showed GFP and CD31 double-positive cells (i.e., ASCs-derived endothelial cells). The mRNA expression of CXCR4 and SDF-1α was much higher in the CXCR4-transfected HBASCs transplants than in the other three transplants. Conclusions: Our data demonstrated that HBASCs can enhance the survival and quality of transplanted free fat tissues. Moreover, CXCR4 transfection of these HBASCs could augment this effect. Stimulation of angiogenesis and decreased fat cell apoptosis due to the recruitment of endothelial progenitor cells (EPCs) and an increase in graft revascularization are potential mechanisms underlying the improved long-term survival of free fat transplants following CXCR4-transfected HBASCs treatment.