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Influence of cryopreserved olfactory ensheathing cells transplantation on axonal regeneration in spinal cord ofadult rats
作者姓名:沈慧勇  殷德振  唐勇  吴燕峰  程志安  杨睿  黄霖
作者单位:[1]DepartmentofOrthopaedics,MemorialHospital,SunYat-senUniversityofMedicalScience,Guangzhou510120,China [2]LingnanUmbilicalBloodBank,SunYat-senUniversityofMedicalScienee,Guangzhou510120,China
摘    要:Objective: To observe the effects of cryopreserved olfactory ensheathing cells (OECs) transplantation on axonal regeneration and functional recovery following spinal cord injury in adult rats. Methods : Twenty-four rats were divided into experimental and control groups, each group having 12 rats. The spinal cord injury was established by transecting the spinal cord at T10 level with microsurgery scissors. OECs were purified from SD rat olfactory bulb and cultured in DMEM ( Dulbecco‘s minimum essential medium) and cryopreserved (-120~C) for two weeks. OECs suspension I (1-1.4) x 105/ul ] was transplanted into transected spinal cord, while the DMEM solution was injected instead in the control group. At 6 and 12 weeks after transplantation, the rats were evaluated with climbing test and MEP ( moter evoked potentials) monitoring. The samples of spinal cord were procured and studied with histological and immunohisto chemical stainings. Results: At 6 weeks after transplantation, all of the rats in both transplanted and control groups were paraplegic, and MEPs could not be recorded. Morphologyof transplanted OECs was normal, and OECs wereinterfused with host well. Axons could regrow into gap tissue between the spinal cords. Both OECs and regrown axons were immunoreactive for MBP. No regrown axons were found in the control group. At 12 weeks after transplantation, 2 rats (2/7) had lower extremities muscle contraction, 2 rats (2/7) had hip and/or knee active movement, and MEP of 5 rats (5/7) could be recorded in the calf in the transplantation group. None of the rats (7/7) in the control group had functional improvement, and none had MEPs recorded. In the transplanted group,histological and immunohistochemical methods showed the number of transplanted OECs reduced and some regrown axons had reached the end of transected spinal cord. However, no regrown axons could be seen except scar formation in the control group. Conclusions: Cryopreserved OECs could integrated with the host and promote regrowing axons across the transected spinal cord ends.

关 键 词:脊髓损伤  轴突  神经再生  细胞移植  成年大鼠  嗅觉鞘细胞  低温保存

Influence of cryopreserved olfactory ensheathing cells transplantation on axonal regeneration in spinal cord of adult rats
SHEN Hui-yong,YIN De-zhen,TANG Yong,WU Yan-feng,CHENG Zhi-an YANG Rui,and HUANG Lin.Influence of cryopreserved olfactory ensheathing cells transplantation on axonal regeneration in spinal cord ofadult rats[J].Chinese Journal of Traumatology(English Edition),2004,7(3):179-183.
Authors:SHEN Hui-yong  YIN De-zhen  TANG Yong  WU Yan-feng  CHENG Zhi-an YANG Rui  and HUANG Lin
Institution:1. Department of Orthopaedics, Memorial Hospital, Sun Yat-sen University of Medical Science, Guangzhou 510120, China
2. Lingnan Umbilical Blood Bank, Sun Yat-sen University of Medical Science, Guangzhou 510120, China
Abstract:OBJECTIVE: To observe the effects of cryopreserved olfactory ensheathing cells (OECs) transplantation on axonal regeneration and functional recovery following spinal cord injury in adult rats. METHODS: Twenty-four rats were divided into experimental and control groups, each group having 12 rats. The spinal cord injury was established by transecting the spinal cord at T10 level with microsurgery scissors. OECs were purified from SD rat olfactory bulb and cultured in DMEM (Dulbecco's minimum essential medium) and cryopreserved (-120 degree) for two weeks. OECs suspension (1-1.4)x10(5)/ul] was transplanted into transected spinal cord, while the DMEM solution was injected instead in the control group. At 6 and 12 weeks after transplantation, the rats were evaluated with climbing test and MEP (moter evoked potentials) monitoring. The samples of spinal cord were procured and studied with histological and immunohistochemical stainings. RESULTS: At 6 weeks after transplantation, all of the rats in both transplanted and control groups were paraplegic, and MEPs could not be recorded. Morphology of transplanted OECs was normal, and OECs were interfused with host well. Axons could regrow into gap tissue between the spinal cords. Both OECs and regrown axons were immunoreactive for MBP. No regrown axons were found in the control group. At 12 weeks after transplantation, 2 rats (2/7) had lower extremities muscle contraction, 2 rats (2/7) had hip and/or knee active movement, and MEP of 5 rats (5/7) could be recorded in the calf in the transplantation group. None of the rats (7/7) in the control group had functional improvement, and none had MEPs recorded. In the transplanted group, histological and immunohistochemical methods showed the number of transplanted OECs reduced and some regrown axons had reached the end of transected spinal cord. However, no regrown axons could be seen except scar formation in the control group. CONCLUSIONS: Cryopreserved OECs could integrated with the host and promote regrowing axons across the transected spinal cord ends.
Keywords:Olfactory nerve  Spinal cord injuries  Axons  Regeneration  Hypothermia
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