Transplantation of human umbilical cord derived-mesenchymal stem cells by different ways for the treatment of spinal cord injury

doi:10.3969/j.issn.2095-4344.2015.14.011

Abstract

Abstract:

BACKGROUND: Currently, human umbilical cord derived-mesenchymal stem cells are mainly for local transplantation, which has some shortcomings, such as large trauma, bleeding, complications, that limit its widespread application in clinical practice.

OBJECTIVE: To investigate the feasibility of intravenous transplantation of human umbilical cord derived-mesenchymal stem cells for repair of spinal cord injury.

METHODS: Eighty Wistar rats with spinal cord hitting were divided into five groups: blank control group with no transplantation (n=10), DMEM local transplantation group (n=15), DMEM intravenous transplantation group (n=15), cell local transplantation group (n=20), cell intravenous transplantation group (n=20). The functional recovery of spinal cord injury was observed with Basso, Beattie and Bresnahan scores at regular time as well as hematoxylin-eosin staining and immunohistochemistry staining.

RESULTS AND CONCLUSION: During 1 day to 2 weeks after transplantation, there was no significant difference in the Basso, Beattie and Bresnahan scores between the five groups; within 4-12 weeks after transplantation, the Basso, Beattie and Bresnahan scores were significantly higher in the two cell transplantation groups than the other three groups, but there was no difference between these two cell transplantation groups (P > 0.05). Histological observation showed that the number of voids and glial scars was less in the cell local transplantation group and cell intravenous transplantation group compared with the other three groups, and there was also no difference between the two cell transplantation groups. These results indicate that the intravenous transplantation of human umbilical cord derived-mesenchymal stem cells is similar to the local transplantation in the repair of acute spinal cord injury, which is simple and avoids secondary injuries and various complications. It is recommended that this method provide a new approach for cell transplantation.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Umbilical Cord, Mesenchymal Stem Cell Transplantation, Injections, Intravenous, Spinal Cord Injuries

CLC Number:

R394.2

Liu Yu-liang, Li Jun, He Yu-qin, Zhuo Feng, Wei Kai-bin. Transplantation of human umbilical cord derived-mesenchymal stem cells by different ways for the treatment of spinal cord injury

[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(14): 2192-2198.

Figures/Tables 3

2.1 人脐带间充质干细胞的培养和鉴定倒置显微镜下观察组织块，于10 d左右见多数组织块周围长出大量新生细胞，形态多为长梭样、纺锤形，主要围绕组织块散在分布(图1)，12-15 d时可见以组织块为中心周围细胞明显增多，细胞形态细瘦，类似成纤维样细胞，大体呈漩涡状或放射状分布。传代培养后的细胞形态上大体一致，第5代时镜下可见大小均一，形态规则的成纤维样细胞(图2)。第3代细胞经过流式鉴定表明CD44、CD29、CD90、CD54阳性表达，而CD31、CD45、CD34、HLA-DR阴性表达，与其他研究结果相同[1-7]。 2.2 实验动物数量分析实验过程中，空白对照组死亡0只，DMEM局部移植组死亡2只，DMEM静脉移植组死亡1只，细胞局部移植组死亡2只，细胞静脉移植组死亡1只，死亡后给予重新补充观察。 2.3 各组大鼠BBB评分见表1，图3。术前所有大鼠双后肢肌力正常，即BBB评分均为21分，造模成功后均呈现双后肢全瘫，移植后第1天BBB评分为0分或1分，随后各组BBB评分呈现逐渐上升的趋势。移植后3周细胞局部移植组、细胞静脉移植组BBB评分分别为7.20±0.70，7.12± 0.81，明显高于空白对照组、DMEM局部移植组、DMEM静脉移植组，差异有显著性意义(P < 0.05)，而细胞局部移植组、细胞静脉移植组间差异无显著性意义(P > 0.05)，空白对照组、DMEM局部移植组、DMEM静脉移植组间差异无显著性意义(P > 0.05)。移植后1 d，1，2周各组之间比较差异无显著性意义(P > 0.05)；移植后4，6，8，10，12周，细胞局部移植组、细胞静脉移植组与其他3组比较差异有显著性意义(P < 0.05)，细胞局部移植组、细胞静脉移植组间差异无显著性意义(P > 0.05)，空白对照组、DMEM局部移植组、DMEM静脉移植组间差异无显著性意义(P > 0.05)。 2.4 苏木精-伊红染色观察结果第8周时苏木精-伊红染色可见空白对照组、DMEM局部移植组、DMEM静脉移植组损伤区域向内凹陷，损伤局部结构紊乱，瘢痕增生，局部形成空洞，细胞走行方向不一致。细胞静脉移植组和细胞局部移植组相近，两组形成的空洞及瘢痕较其他3组都相对减少(图4)。 2.5 免疫荧光染色和免疫组化染色结果移植后4周时，细胞局部移植组和细胞静脉移植组在损伤局部可见BrdU阳性染色的人脐带间充质干细胞，未损伤的部位则未见到明显的阳性染色，阳性细胞数量自损伤部位向两端逐渐越少(图5)。移植后8周时人脐带间充质干细胞大部分集中在损伤的脊髓实质周围，12周时细胞局部移植组和细胞静脉移植组脊髓损伤局部仍然有BrdU阳性染色的人脐带间充质干细胞，但是局部存活的细胞数量比8周时有所减少。空白对照组、DMEM局部移植组和DMEM静脉移植组在8，12周都未见到任何阳性染色细胞。细胞局部移植组和细胞静脉移植组标本在移植后第4周时可见空洞周围主要以纤维性星形胶质细胞为主，第8周时星形胶质细胞数量下降，第12周时脊髓损伤局部形成大量的胶质瘢痕，两组形成的胶质瘢痕面积较其他3组减少。细胞局部移植组、细胞静脉移植组无明显差异，空白对照组、DMEM局部移植组和DMEM静脉移植组之间无明显差异(图6)。"

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