脂肪干细胞与自体富血小板血浆载体复合物体内构建血管化组织工程脂肪

背景：血运重建机制是组织工程化脂肪组织成功构建的决定性因素。目的：观察脂肪干细胞与自体富血小板血浆和纤维蛋白胶载体复合物在体内构建血管化组织工程脂肪的可行性。方法：从健康成年人吸脂术后的脂肪组织中分离脂肪干细胞并行原代及传代培养，将第3代经BrdU标记的脂肪干细胞向脂肪细胞定向诱导2周后，制成浓度为5×1010 L-1细胞悬液。由0.5 mL细胞悬液、100 μL富血小板血浆工作液或DMEM培养基与0.5 mL纤维蛋白胶组成实验组和对照组移植物，分别植入裸鼠背部皮下。结果与结论：植入后8周取材：①实验组可见血管明显增生并长入材料，呈轻度纤维包裹；对照组有少量血管长入材料中，也有轻度纤维包裹现象。实验组新生组织湿质量大于对照组(P < 0.01)。②苏木精-伊红染色均可见移植物中有新生脂肪组织形成和不同程度的微血管长入；实验组微血管数多于对照组(P < 0.01)。③新生组织免疫荧光染色示，两组新生脂肪细胞的胞核及部分微血管内皮细胞的胞核呈现绿色荧光。说明脂肪干细胞与自体富血小板血浆和纤维蛋白胶载体复合物在体内可构建血管化组织工程脂肪，脂肪干细胞与自体富血小板血浆共同参与新生脂肪组织的血管化过程，自体富血小板血浆能促进组织工程构建物的血运重建，保证更多种子细胞的成活。

Construction of vascularized adipose using adipose-derived stem cells and autogeneic platelet-rich plasma carrier complex in vivo by tissue engineering

BACKGROUND:Revascularization mechanism is a determinal factor of successful construction of adipose tissue by tissue engineering.OBJECTIVE:To investigate the feasibility of construction of vasuclarized adipose using adipose-derived stem cells and autogeneic platelet-rich plasma carrier complex in vivo by tissue engineering。METHODS:Adipose-derived stem cells were isolated from the subcutaneous adipose tissue of healthy adult after liposuction, and primary culture and subculture of adipose-derived stem cells were conducted. After being induced towards adipocytes for 2 weeks, 5× 1010/L passage 3 cell suspension labeled by BrdU was prepared. Two groups were included: experimental group, in which 0.5 mL cell suspension, 100 μL platelet-rich plasma and 0.5 mL fibrin glue were implanted into the subcutaneous fascia of nude mice; control group in which 0.5 mL cell suspension, 100 μL DMEM and 0.5 mL fibrin glue were implanted into the subcutaneous fascia of nude mice.RESULTS AND CONCLUSION:At 8 weeks after surgery, neogenetic vessels grew into the scaffolds and mild fiber encapsulation was observed in the experimental group, while few vessels grew into the scaffolds and mild fiber encapsulation was also observed in the control group. The wet weight of cambium in the experimental group was higher than that in the control group (P < 0.01). Hematoxylin-eosin staining showed the formation of neogenetic adipose tissues and the growth of micrangium in the implant. The number of micro vessels in the experimental group was greater than that in the control group (P < 0.01). The immunofluorescence staining of cambium showed that the cell nucleus of regenerated adipocytes and partial capillary endothelium in both groups presented green fluorescence. It is feasible to prepare vasuclarized adipose using adipose-derived stem cells and autogeneic platelet-rich plasma carrier complex in vivo by tissue engineering. Adipose-derived stem cells and autogeneic platelet-rich plasma participate in neovascularization of neogenetic adipose tissue. The autogeneic platelet-rich plasma can promote revascularization of tissue-engineered complex and ensure survival of more seed cells.