Ma Tian, Luan Shao-Liang, Huang Hong, Sun Xing-Kun, Yang Yan-Mei, Zhang Hui, Han Wei-Dong, Li Hong, Han Yan
Department of Plastic and Reconstruction, Chinese PLA General Hospital, Beijing, China (mainland).
Department of Vascular and Endovascular, Chinese PLA General Hospital, Beijing, China (mainland).
Med Sci Monit. 2016 Dec 30;22:5206-5217. doi: 10.12659/msm.902690.
BACKGROUND CC chemokine receptor 7 (CCR7) expression is vital for cell migration to secondary lymphoid organs (SLOs). Our previous work showed that inducing CCR7 expression enabled syngeneic mesenchymal stem cells (MSCs) to migrate into SLOs, resulting in enhanced immunosuppressive performance in mice. Given that human adipose-derived stem cells (hASCs) are widely used in clinical therapy, we further investigated whether upregulation of CCR7 enables xenogeneic hASCs to migrate to rat SLOs. MATERIAL AND METHODS hASCs rarely express CCR7; therefore, hASCs were transfected with lentivirus encoding rat CCR7 (rCCR7) plus green fluorescence protein (GFP) or GFP alone. CCR7 mRNA and cell surface expression of rCCR7-hASCs and GFP-hASCs were examined by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM), respectively. The phenotype, differentiation, and proliferation capacity of each cell type was also determined. To examine migration, rCCR7-hASCs and GFP-hASCs were injected intravenously into Lewis rats, and the proportion of GFP-positive cells in the spleen and lymph nodes was determined with FCM. RESULTS mRNA and cell surface protein expression of CCR7 was essentially undetectable in hASCs and GFP-ASCs; however, CCR7 was highly expressed in rCCR7-ASCs. rCCR7-hASCs, GFP-hASCs, and hASCs shared a similar immunophenotype, and maintained the ability of multilineage differentiation and proliferation. In addition, the average proportion of GFP-positive cells was significantly higher following transplantation of rCCR7-hASCs compared with GFP-hASCs (p<0.01). CONCLUSIONS These results suggest that upregulation of rat CCR7 expression does not change the phenotype, differentiation, or proliferation capacity of hASCs, but does enable efficient migration of hASCs to rat SLOs.
背景 C-C 趋化因子受体 7(CCR7)的表达对于细胞迁移至次级淋巴器官(SLOs)至关重要。我们之前的研究表明,诱导 CCR7 表达可使同基因间充质干细胞(MSCs)迁移至 SLOs,从而增强小鼠的免疫抑制性能。鉴于人脂肪来源干细胞(hASCs)在临床治疗中广泛应用,我们进一步研究了 CCR7 的上调是否能使异种 hASCs 迁移至大鼠 SLOs。
材料与方法 hASCs 很少表达 CCR7;因此,用编码大鼠 CCR7(rCCR7)加绿色荧光蛋白(GFP)的慢病毒或仅 GFP 转染 hASCs。分别通过逆转录-聚合酶链反应(RT-PCR)和流式细胞术(FCM)检测 rCCR7-hASCs 和 GFP-hASCs 的 CCR7 mRNA 及细胞表面 rCCR7 的表达。还测定了每种细胞类型的表型、分化和增殖能力。为检测迁移情况,将 rCCR7-hASCs 和 GFP-hASCs 静脉注射到 Lewis 大鼠体内,并用 FCM 测定脾脏和淋巴结中 GFP 阳性细胞的比例。
结果 在 hASCs 和 GFP-ASCs 中基本检测不到 CCR7 的 mRNA 和细胞表面蛋白表达;然而,CCR7 在 rCCR7-ASCs 中高表达。rCCR7-hASCs、GFP-hASCs 和 hASCs 具有相似的免疫表型,并保持多向分化和增殖能力。此外,与 GFP-hASCs 相比,rCCR7-hASCs 移植后 GFP 阳性细胞的平均比例显著更高(p<0.01)。
结论 这些结果表明,大鼠 CCR7 表达的上调不会改变 hASCs 的表型、分化或增殖能力,但能使 hASCs 有效迁移至大鼠 SLOs。