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[大鼠脐带间充质干细胞修复脊髓损伤]

[Repair of spinal cord injury with rats' umbilical cord MSCs].

作者信息

Zhu Yuhai, Feng Shiqing, Wang Xue

机构信息

Department of Cerebral Surgery, the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, P.R. China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2009 Dec;23(12):1491-6.

Abstract

OBJECTIVE

To study the growth characteristics of umbilical cord MSCs (UCMSCs) in vitro and its effect on the nerve regeneration after spinal cord injury (SCI).

METHODS

UCMSCs isolated from pregnant rats umbilical cord were cultured and purified in vitro. Sixty female Wistar rats weighing (300 +/- 10) g were randomized into three groups (n=20 per group). UCMSCs group (group A) in which UCMSCs suspension injection was conducted; DMEM control group (group B) in which 10% DMEM injection was conducted; sham group (group C) in which the animal received laminectomy only. Establish acute SCI model (T10) by Impactor model-II device in group A and group B. The recovery of the lower extremity was observed using BBB locomotor scoring system, neurofilament 200 (NF-200) immunofluorescence staining was performed to detect the neural regeneration, and then the corticospinal tract (CST) was observed using the biotinylated dextran amine (BDA) tracing.

RESULTS

Cultured UCMSCs were spindle-shaped fibrocyte-like adherent growth, swirling or parallelly. The USMSCs expressed CD29, but not CD31, CD45, and HLA-DR. The BBB score was higher in group A than group B 4, 5, and 6 weeks after operation, and there was a significant difference between two groups (P < 0.05). The BBB scores at different time points were significantly lower in groups A and B than that in group C (P < 0.05). UCMSCs was proved to survive and assemble around the injured place by frozen section of the cords 6 weeks after injury. NF-200 positive response area in groups A, B, and C was (11,943 +/- 856), (7,986 +/- 627), and (13,117 +/- 945) pixels, respectively, suggesting there was a significant difference between groups A, C and group B (P < 0.05), and no significant difference was evident between group A and group C (P > 0.05). BDA anterograde tracing 10 weeks after operation demonstrated that more regenerated nerve fibers went through injured area in group A, but just quite few nerve fibers in group B went through the injuried cavity. The ratios of regenerative axons amount to T5 axons in group A and group B were smaller than that of group C (P < 0.05).

CONCLUSION

UCMSCs can proliferate rapidly in vitro, survive and differentiate to neurons after being grafted into injured spinal cord. The transplantation of UCMSCs is effective in promoting functional recovery and axonal regeneration after SCI.

摘要

目的

研究脐带间充质干细胞(UCMSCs)的体外生长特性及其对脊髓损伤(SCI)后神经再生的影响。

方法

从孕鼠脐带中分离出UCMSCs并在体外进行培养和纯化。将60只体重为(300±10)g的雌性Wistar大鼠随机分为三组(每组n = 20)。UCMSCs组(A组)进行UCMSCs悬液注射;DMEM对照组(B组)注射10% DMEM;假手术组(C组)仅行椎板切除术。A组和B组采用Impactor-II型装置建立急性SCI模型(T10)。采用BBB运动评分系统观察下肢恢复情况,进行神经丝200(NF-200)免疫荧光染色检测神经再生,然后采用生物素化葡聚糖胺(BDA)示踪观察皮质脊髓束(CST)。

结果

培养的UCMSCs呈纺锤形纤维细胞样贴壁生长,呈漩涡状或平行排列。UCMSCs表达CD29,但不表达CD31、CD45和HLA-DR。术后4、5、6周A组的BBB评分高于B组,两组间差异有统计学意义(P < 0.05)。A组和B组不同时间点的BBB评分均显著低于C组(P < 0.05)。损伤后6周通过脊髓冰冻切片证实UCMSCs存活并聚集在损伤部位周围。A组、B组和C组的NF-200阳性反应面积分别为(11,943±856)、(7,986±627)和(13,117±945)像素,提示A组、C组与B组之间差异有统计学意义(P < 0.05),A组与C组之间差异无统计学意义(P > 0.05)。术后10周BDA顺行示踪显示,A组有更多再生神经纤维穿过损伤区域,而B组仅有极少数神经纤维穿过损伤腔。A组和B组再生轴突数量与T5轴突数量的比值均小于C组(P < 0.05)。

结论

UCMSCs在体外可快速增殖,移植到损伤脊髓后可存活并分化为神经元。UCMSCs移植对促进SCI后的功能恢复和轴突再生有效。

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