神经干细胞与脊髓脱细胞支架体外共培养的可行性

doi:10.3969/j.issn.1006-9771.2021.01.011

《中国康复理论与实践》 ›› 2021, Vol. 27 ›› Issue (1): 71-78.doi: 10.3969/j.issn.1006-9771.2021.01.011

神经干细胞与脊髓脱细胞支架体外共培养的可行性

漆国栋^1,2, 江琼², 伍亚民³, 申开琴¹, 杨琴², 漆伟¹

1.重庆市中医骨科医院骨科，重庆市 400010
2.重庆医科大学中医药防治代谢性疾病重庆市重点实验室，重庆市 400016
3.陆军军医大学大坪医院野战外科研究所，创伤烧伤与复合伤国家重点实验室，重庆市 400042

收稿日期:2020-03-10 修回日期:2020-04-02 出版日期:2021-01-25 发布日期:2021-01-27
通讯作者: 漆伟，E-mail： qiwei200@126.com
作者简介:漆国栋(1993-)，男，汉族，重庆市人，硕士，医师，主要研究方向：骨与神经组织损伤后修复方面的研究；漆伟(1966-)，男，汉族，重庆市人，硕士，主任医师，主要研究方向：骨与神经组织损伤后修复方面的研究。
基金资助:
1. 重庆市卫健委中医药科技项目(No. ZY201702134);2. 重庆市科卫联合科技项目(No. 2019ZY023495;No.2019ZY023292)

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

QI Guo-dong^1,2, JIANG Qiong², WU Ya-min³, SHEN Kai-qin¹, YANG Qin², QI Wei¹

1. Orthopedic Department, Chongqing Orthopedic Hospital of Traditional Chinese Medicine, Chongqing 400010, China
2. Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, Chongqing Medical University, Chongqing 400016, China
3. Field Surgery Research Institute, Daping Hospital, PLA Military Medical University, State Key Laboratory of Trauma Burn and Complex Injury, Chongqing 400042, China

Received:2020-03-10 Revised:2020-04-02 Published:2021-01-25 Online:2021-01-27
Contact: QI Wei, E-mail: qiwei200@126.com
Supported by:
Chongqing Health and Family Planning Commission Traditional Chinese Medicine Research Project (No. ZY201702134) and Chongqing Health and Family Planning Commission and Science and Technology Bureau Research Project (No. 2019ZY023495 and No. 2019ZY023292)

摘要/Abstract

摘要： 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。

关键词: 脊髓损伤, 神经干细胞, 脊髓脱细胞支架, 体外共培养, 脊髓组织工程

Abstract: 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.

Key words: spinal cord injury, neural stem cells, spinal cord acellular scaffold, co-culture in vitro, spinal cord tissue engineering

中图分类号:

R318.5

漆国栋, 江琼, 伍亚民, 申开琴, 杨琴, 漆伟. 神经干细胞与脊髓脱细胞支架体外共培养的可行性[J]. 《中国康复理论与实践》, 2021, 27(1): 71-78.

QI Guo-dong, JIANG Qiong, WU Ya-min, SHEN Kai-qin, YANG Qin, QI Wei. Co-culture of Neural Stem Cells and Spinal Cord Acellular Scaffold in Vitro[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2021, 27(1): 71-78.

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神经干细胞与脊髓脱细胞支架体外共培养的可行性

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

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