诱导人眼轮匝肌来源肌源干细胞向许旺细胞样细胞的分化

背景:肌源干细胞的优越性引导学者们尝试从人眼轮匝肌中分离该细胞,同时进行许旺细胞方向的诱导分化,为周围神经的修复提供新的种子细胞来源。目的:诱导人肌源干细胞向具有许旺细胞特性的细胞分化。方法:①收集重睑成形术中切除的上睑眼轮匝肌,经酶消化和细胞筛过滤,贴壁培养分离人肌源干细胞,予细胞特异标记物以免疫组织化学染色。②取大鼠坐骨神经分离培养许旺细胞,收集许旺细胞条件培养液。③将人肌源干细胞与许旺细胞条件培养液共培养,观察转化细胞的形态和免疫组织化学染色的变化。结果与结论:①原代培养4周时可见人肌源干细胞,与传代培养之人肌源干细胞同样呈现明显的小圆形形态,折光性强,少量细胞呈现短梭形。所有人肌源干细胞表现为 Desmin 阳性,Sca-1 染色阳性。②分离培养大鼠坐骨神经来源许旺细胞S100染色阳性率为(97.4±0.7)%。③经与许旺细胞条件培养液共培养,人肌源干细胞分化后细胞表达许旺细胞特异标记物S100,GFAP和p75。结果提示分离获得人肌源干细胞与许旺细胞条件培养液共培养,可以诱导人肌源干细胞表达许旺细胞特异标记物,初步证实了人肌源干细胞可分化为许旺细胞样细胞。

Differentiation potential of human Orbicularis oculi muscle-derived stem cells towards Schwann cells phenotype

BACKGROUND: Muscle derived stem cells (MDSCs) can be isolated from human orbicularis oculi muscle and be differentiated to a Schwann cell phenotype which could eventually provide functional benefits for peripheral nerve repair. OBJECTIVE: To induce the differentiation of MDSCs into Schwann cell phenotype. METHODS: ①Under the support of microscope, we collected the discarded human Orbicularis oculi muscle resected in the upper eyelid blepharoplasty and isolate human-MDSCs within it with aid of tri-enzyme digestion and pre-plating technique, and then identify the cells by immunohistochemistry method. ②We isolated Schwann cells and identify the cells by immunohistochemistry method. Through half-harvest method, we would like to prepare conditioned medium from Schwann cell culture. ③We co-culture human-MDSCs with Schwann cell conditioned medium and the transdifferentiated cell morphology was investigated daily under microscope. The common used marker, S-100, GFAP and p75 were stained to identify Schwann cell phenotype with use of immunohistochemistry method. RESULTS AND CONCLUSION: ①We collected human Orbicularis oculi muscle sample from three young female volunteer with their consensus. Human-MDSCs were isolated from Orbicularis oculi muscle and have their desmin positively stained and Sca-1 was positively expressed. ②Schwann cells were isolated and identified with S-100 positively stained at the rate of (97.4±0.7)%. ③The isolated human-MDSCs were successfully transdifferentiated into Schwann cell-like cells with positive expression of S100, GFAP and p75, which would serve as a unanimous evidence of Schwann cell phenotype. Human-MDSCs could be transdifferentiated into Schwann cell-like cells when co-cultured within Schwann cell conditioned medium, which would serve as an alternative candidate for commonly studied Schwann cells in tissue engineering nerve graft.