中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
23期
4555-4558
,共4页
脊髓损伤%干细胞移植%脑源性神经营养因子
脊髓損傷%榦細胞移植%腦源性神經營養因子
척수손상%간세포이식%뇌원성신경영양인자
MSCs transplantation upregulatas the expression of BDNF%which promotes neurite regeneration. It may be one of the mechanisms of repairing SCl
背景:近年来关于骨髓间充质干细胞移植对脊髓损伤修复方面的研究较多,但目前相关机制尚不清楚.目的:观察间充质干细胞移植对大鼠脊髓损伤后脑源性神经营养因子表达的影响.设计、时间及地点:随机对照动物实验,于2003-04/07在中国医科大学神经解剖学实验室完成.材料:选取鼠龄3个月的SD大鼠64只,雌雄不拘,体质量250～300 g,随机抽取4只大鼠用于骨髓间充质干细胞的分离与培养,其余60只用于制备脊髓横断损伤模型.方法:60只大鼠随机抽签法分为3组,细胞移植组(n=24):脊髓损伤后第7天,无菌条件下以微量注射缓慢注入含骨髓间充质干细胞(1×109L-1)的培养液5μL至脊髓损伤区;PBS组(n=24):注入等量(5μL)磷酸盐缓冲液体;空白对照组(n=12):注入生理盐水5μL.主要观察指标:分别于造模后7,14,28 d取材,观察骨髓间充质干细胞的形态变化;免疫组织化学法检测间充质干细胞移植后大鼠脊髓损伤区脑源性神经营养因子的表达.结果:60只SD大鼠均进入结果分析.10代以后细胞增殖能力有所减弱,胞体变得扁平,若加入碱性成纤维细胞生长因子,则可维持其增殖能力和形态.大鼠脑源性神经营养因子在正常大鼠脊髓组织中有一定表达,细胞移植后第7,14,28天,细胞移植组脑源性神经营养因子表达均高于PBS组和空白对照组(P＜0.05);空白对照组与PBS组脑源性神经营养因子表达无明显差异(P＞0.05).结论:骨髓间充质干细胞在移植后通过上调脑源性神经营养因子的表达从而促进轴突的再生,可能是治疗脊髓损伤的重要机制.
揹景:近年來關于骨髓間充質榦細胞移植對脊髓損傷脩複方麵的研究較多,但目前相關機製尚不清楚.目的:觀察間充質榦細胞移植對大鼠脊髓損傷後腦源性神經營養因子錶達的影響.設計、時間及地點:隨機對照動物實驗,于2003-04/07在中國醫科大學神經解剖學實驗室完成.材料:選取鼠齡3箇月的SD大鼠64隻,雌雄不拘,體質量250～300 g,隨機抽取4隻大鼠用于骨髓間充質榦細胞的分離與培養,其餘60隻用于製備脊髓橫斷損傷模型.方法:60隻大鼠隨機抽籤法分為3組,細胞移植組(n=24):脊髓損傷後第7天,無菌條件下以微量註射緩慢註入含骨髓間充質榦細胞(1×109L-1)的培養液5μL至脊髓損傷區;PBS組(n=24):註入等量(5μL)燐痠鹽緩遲液體;空白對照組(n=12):註入生理鹽水5μL.主要觀察指標:分彆于造模後7,14,28 d取材,觀察骨髓間充質榦細胞的形態變化;免疫組織化學法檢測間充質榦細胞移植後大鼠脊髓損傷區腦源性神經營養因子的錶達.結果:60隻SD大鼠均進入結果分析.10代以後細胞增殖能力有所減弱,胞體變得扁平,若加入堿性成纖維細胞生長因子,則可維持其增殖能力和形態.大鼠腦源性神經營養因子在正常大鼠脊髓組織中有一定錶達,細胞移植後第7,14,28天,細胞移植組腦源性神經營養因子錶達均高于PBS組和空白對照組(P＜0.05);空白對照組與PBS組腦源性神經營養因子錶達無明顯差異(P＞0.05).結論:骨髓間充質榦細胞在移植後通過上調腦源性神經營養因子的錶達從而促進軸突的再生,可能是治療脊髓損傷的重要機製.
배경:근년래관우골수간충질간세포이식대척수손상수복방면적연구교다,단목전상관궤제상불청초.목적:관찰간충질간세포이식대대서척수손상후뇌원성신경영양인자표체적영향.설계、시간급지점:수궤대조동물실험,우2003-04/07재중국의과대학신경해부학실험실완성.재료:선취서령3개월적SD대서64지,자웅불구,체질량250～300 g,수궤추취4지대서용우골수간충질간세포적분리여배양,기여60지용우제비척수횡단손상모형.방법:60지대서수궤추첨법분위3조,세포이식조(n=24):척수손상후제7천,무균조건하이미량주사완만주입함골수간충질간세포(1×109L-1)적배양액5μL지척수손상구;PBS조(n=24):주입등량(5μL)린산염완충액체;공백대조조(n=12):주입생리염수5μL.주요관찰지표:분별우조모후7,14,28 d취재,관찰골수간충질간세포적형태변화;면역조직화학법검측간충질간세포이식후대서척수손상구뇌원성신경영양인자적표체.결과:60지SD대서균진입결과분석.10대이후세포증식능력유소감약,포체변득편평,약가입감성성섬유세포생장인자,칙가유지기증식능력화형태.대서뇌원성신경영양인자재정상대서척수조직중유일정표체,세포이식후제7,14,28천,세포이식조뇌원성신경영양인자표체균고우PBS조화공백대조조(P＜0.05);공백대조조여PBS조뇌원성신경영양인자표체무명현차이(P＞0.05).결론:골수간충질간세포재이식후통과상조뇌원성신경영양인자적표체종이촉진축돌적재생,가능시치료척수손상적중요궤제.
BACKGROUND: Transplantation of bone marrow mesenchymal stem cells (MSCs) improves functional recovery after spinal cord injury (SCl), but the mechanisms involved remain unclear.OBJECTIVE: To observe the effects of MSCs transplantation on the expression of brain-derived neurotrophic factors (BDNF) in rats after SCl.DESIGN, TIME AND SETTING: Randomized, controlled, animal experiment. The study was performed at the Laboratory of Neuroanatomy, China Medical University from April to July 2003.MATERIALS: A total of 64 SD rats, aged 3 months, of either gender, weighing 250-300 g, were used. Of them, 4 were randomly selected to isolate and culture MSCs, and the remaining were used to establish SCl models.METHODS: The 60 rats were randomly divided into 3 groups. Seven days after SCl, MSCs group (n=24) was transplanted with 5 μL culture solution containing 1×109/L MSCs to the injury site using micro-injection; PBS group (n=24) was transplanted with 5μ L PBS, and the blank control group (n=12) with 5μ L normal saline.MAIN OUTCOME MEASURES: The rats were sacrificed at 7, 14, and 28 days post-surgery. MSCs morphology was observed and the expression of BDNF at the lesion areas was examined by immunohistochemistry,RESULTS: All 60 rats were included in the final analysis. After ten subcultures, the cell proliferative capacity was reduced, and cell body turned to flat; the MSCs protiferation and morphous could be maintained by adding basic fibroblast growth factor.Transplantation of MSCs enhanced the expression of BDNF compared with PBS and blank control groups at 7, 14, and 28 days post-surgery (P ＜ 0.05); white no significant difference was found between PBS and blank control groups (P ＞ 0.05).
