神经干细胞与脊髓脱细胞支架体外共培养的可行性

目的 观察大鼠神经干细胞(NSCs)在脊髓脱细胞支架(SCAS)上黏附、生长和分化情况，评价其构建脊髓组织工程的可行性。 方法 新生Sprague-Dawley大鼠大脑皮质来源NSCs传代培养并鉴定；Sprague-Dawley雌性大鼠脊髓组织改良物理振荡结合化学萃取法制备SCAS，并行基础评估；将第三代NSCs种植在SCAS上，体外共培养，免疫荧光、免疫组化和扫描电镜观察支架上细胞形态。 结果 共培养的细胞为能增殖、分化的NSCs。制备的SCAS孔隙率、含水率、酶解率都明显高于正常脊髓(|t| > 4.679, P < 0.01)；SCAS基质结构呈疏松网络状，基质上可见极少量残留细胞核。NSCs在SCAS上黏附、生长良好，可分化为神经元和神经胶质细胞。 结论 制备的SCAS脱细胞较彻底，具有多通道空间结构，适合NSCs黏附、生长和分化，可用于脊髓组织工程。

Co-culture of Neural Stem Cells and Spinal Cord Acellular Scaffold in Vitro

Objective To observe the adhesion, growth and differentiation of rat neural stem cells (NSCs) on spinal cord acellular scaffold (SCAS) to evaluate its feasibility for spinal cord tissue engineering. Methods NSCs derived from neonatal Sprague-Dawley rat cerebral cortex were cultured and identified. SCAS were prepared from female Sprague-Dawley rat spinal cord tissues using modified chemical extraction and physical oscillation, and evaluated. The third generation NSCs were planted on SCAS and co-cultured, the morphology of the cells on the scaffold was observed with immunofluorescence, immunohistochemistry and scanning electron microscope. Results The cultured cells were NSCs, which could proliferate and differentiate. The porosity, water content and enzymatic hydrolysis rates of the prepared SCAS were significantly higher than that of normal spinal cord (|t| > 4.679, P < 0.01). The matrix structure of SCAS was loosely network-like, with few residual nuclei. NSCs adhered and grew well, and differentiated into neurons and glial cells on SCAS. Conclusion This kind of SCAS shapes multi-channel spatial structure and is suitable for NSCs adhesion, growth and differentiation, which can be used for spinal cord tissue engineering.