中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (45): 8424-8429.doi: 10.3969/j.issn.2095-4344.2012.45.012

• 脂肪干细胞 adipose-derived stem cells • 上一篇    下一篇

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

黎洪棉1，2，柳大烈3，赵培冉4，梁双武4   

  1. 1中山大学附属中山医院博士后科研工作站，整形美容外科，广东省中山市 528403
    2南方医科大学博士后科研流动站，广东省广州市 510515
    3南方医科大学珠江医院整形外科，广东省广州市 510282
    4南方医科大学组织工程研究中心，广东省广州市 510515
  • 收稿日期:2012-01-02 修回日期:2012-01-09 出版日期:2012-11-04 发布日期:2012-11-04
  • 通讯作者: 柳大烈，教授，主任医师，博士(后)导师，南方医科大学珠江医院整形外科，广东省广州市 510282 Liudalie@hotmail.com
  • 作者简介:黎洪棉☆，男，1971年生，广西壮族自治区贺州市人，汉族，2008年南方医科大学毕业，博士后，副主任医师，硕士生导师，主要从事整形美容外科和干细胞、组织工程与再生医学的研究。 binrong2112@163.com

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

Li Hong-mian1, 2, Liu Da-lie3, Zhao Pei-ran4, Liang Shuang-wu4   

  1. 1Postdoctoral Research Center, Department of Plastic and Aesthetic Surgery, the Affiliated Zhongshan Hospital of Sun Yat-Sen University, Zhongshan 528403, Guangdong Province, China
    2Postdoctoral Research Center, Southern Medical University, Guangzhou 510515, Guangdong Province, China
    3Department of Plastic and Reconstructive Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou 501282, Guangdong Province, China
    4Research Center for Tissue Engineering, Southern Medical University, Guangzhou 510515, Guangdong Province, China
  • Received:2012-01-02 Revised:2012-01-09 Online:2012-11-04 Published:2012-11-04
  • Contact: Liu Da-lie, Professor, Chief physician, Doctoral supervisor, Department of Plastic and Reconstructive Surgery, Zhujiang Hospital of Southern Medical University, Guangzhou 501282, Guangdong Province, China Liudalie@hotmail.com
  • About author:Li Hong-mian☆, Doctor, Associate chief physician, Master’s supervisor, Postdoctoral Research Center, Department of Plastic and Aesthetic Surgery, the Affiliated Zhongshan Hospital of Sun Yat-Sen University, Zhongshan 528403, Guangdong Province, China binrong2112@163.com

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490

被引次数

2

摘要：

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。

关键词: 人脂肪干细胞, 自体富血小板血浆, 组织工程, 脂肪形成, 血管发生, 干细胞

Abstract:

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.

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