摘要：
Background: Adipose-derived stem cells (ADSCs) were effective in treating wound. Stromal cell-derived factor-1 (SDF-1), a chemokine usually called CXCL12, is well known for its chemotaxis in induction of cell migration. However, little is known about the SDF-1 responsible for the complex migration of ADSCs from residence to injured sites.
Objective: Herein, we firstly showed SDF-1 is a major regulator involved in migration of ADSCs during wound repair in vivo.
Methods: Trauma in rats was induced by surgical operation. The levels of SDF-1 in wounded tissue were assayed by ELISA. ADSCs were labeled with Green Fluorescent Protein (GFP), and then were transferred to injured rats by intracarotid injection. The plasma levels of ADSCs during wound healing were detected by flow cytometry, and ADSCs in injured tissue were evaluated by bioluminescence imaging in vivo and laser confocal microscopy (LCM), respectively.
Results: ADSCs were successfully labeled with GFP. SDF-1 level reached to the peak value on 24 h after injury and then decreased continuously. Additionally, levels of plasma ADSCs in SDF-1 treated rats reached to the peak value (12%) at d21 after medicine delivery, while those of normal and injured rats showed the peak values of 6.28% and 9.84% at d7 and d21, respectively. Finally, the results of LCM indicated treatment of ectogenic SDF-1 obviously enhanced GFPeADSCs distribution in wounded tissues.
Conclusion: Our results indicated that SDF-1 treatment obviously promoted the migration and directed distribution of ADSCs in traumatic tissue.
关键字:脂肪干细胞ADSCs 、基质细胞衍生因子1、创伤、颈动脉注射
技术流程
研究背景
脂肪干细胞(ADSCs) 在创伤治疗过程中是非常有效的，基质细胞衍生因子1(SDF-1)是一种被经常叫作CXCL12的趋化因子，它因为在细胞迁移感应的趋药性被人们所熟知。然而，很少有人知道SDF-1在创伤组织中ADSCs的作用。
研究结果
1.利用慢病毒转染，GFP成功标记脂肪干细胞（ADSCs），流式细胞术检测成功标记GFP的脂肪干细胞（ADSCs）达到了80%以上
2、创伤修复过程中，ELISA实验检测到受伤组织中基质细胞衍生因子1(SDF-1)的表达增强。SDF-1的表达在24h达到峰值，随后下降。结果表明，在创伤修复过程中，SDF-1是脂肪干细胞（ADSCs）迁移的一种重要的趋化因子。
表1 创伤组织提取液中的SDF-1含量
|
Time after wound healing(h) |
SDF-1 levels (ng/L) |
|
0 6 12 24 48 72 |
0 2.87 ± 0.54 6 3.11 ± 0.61 12 4.24 ± 0.50 24 5.36 ± 0.26 48 4.42 ± 0.30 72 3.74 ± 0.16 |
Data are presented as mean ± S.D. (n =5).
3.外源SDF-1处理的血浆中脂肪干细胞（ADSCs）的迁移增强。流式细胞术检测不同时间点血浆中脂肪干细胞（ADSCs）的水平，在21d时达到峰值（12%），显著高于正常组和未处理组，结果表明，SDF-1处理能显著增强脂肪干细胞（ADSCs）的迁移。
注：流式细胞仪检测血浆中的ADSCs，A：SDF-1处理创伤的大鼠；B未处理创伤的大鼠；C正常大鼠；D血浆中ADSCs的百分比
4.外源SDF-1增强创伤区脂肪干细胞（ADSCs）的迁移。利用生物发光成像检测创伤组织中的GFP-ADSCs，不同处理时间点的结果显示，在SDF-1的存在下，创伤组织中GFP-ADSCs的迁移增强
5.外源SDF-1对创伤区脂肪干细胞（ADSCs）分布的富集。实验处理后不同时间点的创伤组织做冰冻切片，利用共聚焦显微镜进行观察，对比PBS处理组，SDF-1处理组脂肪干细胞（ADSCs）的数量呈现出明显的增长。结果表明：SDF-1增强了脂肪干细胞（ADSCs）的迁移，以此加速创伤修复。
伯信合作技术
慢病毒转染、生物成像、流式细胞检测、共聚焦检测
参考文献
Wu Q, Ji F K, Wang J H, et al. Stromal cell-derived factor 1 promoted migration of adipose-derived stem cells to the wounded area in traumatic rats.[J]. Biochemical & Biophysical Research Communications, 2015, 467(1):140-5.
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