Department of Plastic Surgery, First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.
Department of Pathophysiology, China Medical University, Shenyang, Liaoning 110001, P.R. China.
Mol Med Rep. 2017 Nov;16(5):7297-7304. doi: 10.3892/mmr.2017.7570. Epub 2017 Sep 21.
The ultimate goal of treating peripheral nerve defects is reconstructing continuity of the nerve stumps to regain nerve conduction and functional recovery. Clinically, autologous nerve grafts and Schwann cell (SC) therapy have limitations, such as the need for secondary surgery, sacrifice of another nerve and donor site complication. Adipose derived stem cells (ADSCs) may promise to be ideal alternative cells of SCs. To explore the potential of ADSCs promoting peripheral nerve regeneration, the present study investigated the influences of ADSCs on proliferation and neurotrophic function of SCs using co‑culture model in vitro. Western blot analysis, immunocytochemistry, a cell viability assay, reverse transcription‑polymerase chain reaction (RT‑PCR) and ELISA were applied for examining the interaction of ADSCs and SCs in a co‑culture model in vitro. Western blot analysis and immunocytochemistry demonstrated that protein expression levels of glial filament acidic protein (GFAP) and S100 in ADSCs co‑cultured with SCs for 14 days were significantly higher compared with cells cultured alone. Cell viability assay indicated that the cell viability of SCs co‑cultured with ADSCs for 3, 4, 5, 6 and 7 days was significantly higher than those cultured alone. RT‑PCR showed that expression levels of neurotrophic factors [nerve growth factor (NGF) and glial cell line‑derived neurotrophic factor (GDNF)] and extracellular matrix components [fibronectin (FN) and laminin (LN)] in SCs co‑cultured with ADSCs for 14 days were significantly higher than those in SCs cultured alone. NGF, GDNF, FN and LN in the supernatants of co‑culture system were significantly higher than cells cultured alone, as ELISA revealed. The results of this study suggested that the transplantation of ADSCs may have a promoting potential to the peripheral nerve regeneration as undifferentiated state.
治疗周围神经缺损的最终目标是重建神经残端的连续性，以恢复神经传导和功能恢复。临床上，自体神经移植和施万细胞（SC）治疗存在需要二次手术、牺牲另一根神经和供体部位并发症等局限性。脂肪来源的干细胞（ADSCs）可能有望成为 SC 的理想替代细胞。为了探索 ADSCs 促进周围神经再生的潜力，本研究通过体外共培养模型研究了 ADSCs 对 SC 增殖和神经营养功能的影响。采用 Western blot 分析、免疫细胞化学、细胞活力测定、逆转录-聚合酶链反应（RT-PCR）和 ELISA 检测共培养模型中 ADSCs 与 SC 之间的相互作用。Western blot 分析和免疫细胞化学显示，与单独培养的细胞相比，与 SC 共培养 14 天的 ADSCs 中胶质纤维酸性蛋白（GFAP）和 S100 的蛋白表达水平明显升高。细胞活力测定表明，与单独培养的细胞相比，与 ADSCs 共培养 3、4、5、6 和 7 天的 SC 细胞活力明显升高。RT-PCR 显示，与单独培养的 SC 相比，与 ADSCs 共培养 14 天的神经营养因子[神经生长因子（NGF）和胶质细胞源性神经营养因子（GDNF）]和细胞外基质成分[纤连蛋白（FN）和层粘连蛋白（LN）]的表达水平明显升高。ELISA 显示，共培养系统上清液中的 NGF、GDNF、FN 和 LN 明显高于单独培养的细胞。本研究结果表明，ADSCs 的移植可能具有促进周围神经再生的潜力，因为其处于未分化状态。
