Spinal cord regeneration using human marrow stromal cells

摘要1人骨髓基质细胞是一种具有吸引力的自体细胞和基因治疗的来源。在本研究中，我们开发了一种高效的hMSCs转染方法。尽管非病毒载体往往表现出有效的递送，但它们在基因治疗中提供了优于病毒载体的几个优势。将编码绿色荧光蛋白组分的质粒pEGFP-N1通过核转染和电穿孔两种非病毒方法转染hMSCs和大鼠骨髓基质细胞。核转染法比电穿孔法具有更高的转染率，尤其是在hMSCs中。摘要2我们研究了软骨素酶ABC对轴突再生的影响。我们比较了15 mm胫神经切除和修复后的轴突再生与填充有I型胶原凝胶的硅胶管（阴性对照组），填充有含有三种不同浓度（2.5u/ml，5 u/ml，10 u/ml）的软骨素酶ABC的I型胶原凝胶的硅胶管（软骨素酶ABC组），以及自体神经段（自体神经移植组）。术后12周行神经电生理和组织学检查，除阴性对照组外，其余各组均记录复合动作电位（CMAP）和神经传导速度（NCV）。虽然CMAP和NCV在自体神经移植组中最高，但三个软骨素酶ABC组在任一参数上均无显著差异。组织学结果与电生理结果一致。我们的结论是，局部注射软骨素酶ABC可以显着增加神经管化或人工神经放置的神经缺损修复的临界长度。

Abstract 1Human marrow stromal cells(hMSCs) are an attractive source for autologous cell and gene therapies. In this study, we developed a highly efficient transfection method for hMSCs. Although they tend to show efficient delivery, nonviral vectors offer several advantages over viral vectors for gene therapies. hMSCs and rat marrow stromal cells were transfected with the plasmid pEGFP-N1 that encoded a green fluorescent protein component by using two nonviral methods : nucleofection and electroporation. Nucleofection provided a much better rate of transfer than electroporation particularly in hMSCs.Abstract 2we evaluated the effects of chondroitinase ABC on axonal regeneration across peripheral nerve gaps. We compared axonal regeneration after 15 mm tibial nerve resection and repair with a silicone tube filled with type I collagen gel (negative control group),with a silicone tube filled with type I collagen gel containing chondroitinase ABC at three different concentrations (2.5u/ml, 5u/ml, 10 u/ml) (chondroitinase ABC groups), and with an autologous nerve segment (nerve autograft group). Electrophysiological and histological assessments were carried out 12 weeks after surgery.Compound action potentials (CMAPs) and nerve conduction velocities (NCVs) were recorded in all groups but the negative control group. Although both CMAPs and NCVs were highest in the nerve autograft group, there were no significant differences among the three chondroitinase ABC groups in either parameter. Histological findings were consistent with electrophysiological results. We conclude that topical injection of chondroitinase ABC can significantly increase the critical length of nerve gap repair by tubulization or artificial nerve placement.