中国综合临床
中國綜閤臨床
중국종합림상
CLINICAL MEDICINE OF CHINA
2012年
8期
805-809
,共5页
刘驰%刘杰%郑自龙%蒋伟
劉馳%劉傑%鄭自龍%蔣偉
류치%류걸%정자룡%장위
神经微移植%胚胎纹状体神经元%兴奋毒性纹状体毁损模型%绿色荧光蛋白
神經微移植%胚胎紋狀體神經元%興奮毒性紋狀體燬損模型%綠色熒光蛋白
신경미이식%배태문상체신경원%흥강독성문상체훼손모형%록색형광단백
Micro-transplantation%Embryonic striatal neurons%Exeitotoxic striatal lesion of rmodel%Green fluorescent protein
目的 基于单细胞悬液的应用分别计数由神经微移植(MT)技术及传统移植(TT)技术注入模型纹状体后移植物中DARPP-32阳性细胞,探讨不同效果产生的技术基础与内在机制.方法 移植物由胎龄15 d大鼠胚胎神经节隆起细胞获得并表达绿色荧光蛋白,移植入单侧喹啉酸(QA)毁损大鼠的纹状体内,其中一组以神经微移植设备植入(MT组),另一组则以传统金属针头移植方式植入(TT组),两组分别使用直径为50μm的超薄玻璃毛细管和直径500 μm的金属针头,宿主接受相同的移植细胞总量.结果 移植4个月后行组织学评估,MT组与'TT组具有相似的总移植物体积[(2.8±0.2)mm3与(2.5 ±0.4)mm3,F=0.25,P＞0.05],同时在DARPP-32阳性斑块体积[MT组(0.6±0.1)mm3与TT组(0.6±0.2) mm’,F=0.90,P＞0.05]及酪氨酸羟化酶(TH)染色斑块体积[MT组(1.0 ±0.1)mm3与TT组(0.7±0.1)mm3,F=1.44,P＞0.05]方面具有类似表现.DARPP-32阳性神经元的细胞计数:应用Abercrombie校正公式计算两个移植组的DARPP-32阳性细胞数目,结果分析可见MT组的DARPP-32阳性细胞数(20.1×103±1.2×103)近似于TT组(9.8 ×103±3.2×103)的2.0倍,组间比较差异有统计学意义(F=8.62,P＜0.05),表明更高的DARPP-32阳性细胞计数在MT移植策略中移植物呈现出更好的生长发育结果.结论 神经微移植技术可以增加新生纹状体神经元的数量,其机制可能是移植物-宿主接触面积的增大使得移植物更强地暴露于宿主环境诱导因素之下产生更小的机械损伤.
目的 基于單細胞懸液的應用分彆計數由神經微移植(MT)技術及傳統移植(TT)技術註入模型紋狀體後移植物中DARPP-32暘性細胞,探討不同效果產生的技術基礎與內在機製.方法 移植物由胎齡15 d大鼠胚胎神經節隆起細胞穫得併錶達綠色熒光蛋白,移植入單側喹啉痠(QA)燬損大鼠的紋狀體內,其中一組以神經微移植設備植入(MT組),另一組則以傳統金屬針頭移植方式植入(TT組),兩組分彆使用直徑為50μm的超薄玻璃毛細管和直徑500 μm的金屬針頭,宿主接受相同的移植細胞總量.結果 移植4箇月後行組織學評估,MT組與'TT組具有相似的總移植物體積[(2.8±0.2)mm3與(2.5 ±0.4)mm3,F=0.25,P＞0.05],同時在DARPP-32暘性斑塊體積[MT組(0.6±0.1)mm3與TT組(0.6±0.2) mm’,F=0.90,P＞0.05]及酪氨痠羥化酶(TH)染色斑塊體積[MT組(1.0 ±0.1)mm3與TT組(0.7±0.1)mm3,F=1.44,P＞0.05]方麵具有類似錶現.DARPP-32暘性神經元的細胞計數:應用Abercrombie校正公式計算兩箇移植組的DARPP-32暘性細胞數目,結果分析可見MT組的DARPP-32暘性細胞數(20.1×103±1.2×103)近似于TT組(9.8 ×103±3.2×103)的2.0倍,組間比較差異有統計學意義(F=8.62,P＜0.05),錶明更高的DARPP-32暘性細胞計數在MT移植策略中移植物呈現齣更好的生長髮育結果.結論 神經微移植技術可以增加新生紋狀體神經元的數量,其機製可能是移植物-宿主接觸麵積的增大使得移植物更彊地暴露于宿主環境誘導因素之下產生更小的機械損傷.
목적 기우단세포현액적응용분별계수유신경미이식(MT)기술급전통이식(TT)기술주입모형문상체후이식물중DARPP-32양성세포,탐토불동효과산생적기술기출여내재궤제.방법 이식물유태령15 d대서배태신경절륭기세포획득병표체록색형광단백,이식입단측규람산(QA)훼손대서적문상체내,기중일조이신경미이식설비식입(MT조),령일조칙이전통금속침두이식방식식입(TT조),량조분별사용직경위50μm적초박파리모세관화직경500 μm적금속침두,숙주접수상동적이식세포총량.결과 이식4개월후행조직학평고,MT조여'TT조구유상사적총이식물체적[(2.8±0.2)mm3여(2.5 ±0.4)mm3,F=0.25,P＞0.05],동시재DARPP-32양성반괴체적[MT조(0.6±0.1)mm3여TT조(0.6±0.2) mm’,F=0.90,P＞0.05]급락안산간화매(TH)염색반괴체적[MT조(1.0 ±0.1)mm3여TT조(0.7±0.1)mm3,F=1.44,P＞0.05]방면구유유사표현.DARPP-32양성신경원적세포계수:응용Abercrombie교정공식계산량개이식조적DARPP-32양성세포수목,결과분석가견MT조적DARPP-32양성세포수(20.1×103±1.2×103)근사우TT조(9.8 ×103±3.2×103)적2.0배,조간비교차이유통계학의의(F=8.62,P＜0.05),표명경고적DARPP-32양성세포계수재MT이식책략중이식물정현출경호적생장발육결과.결론 신경미이식기술가이증가신생문상체신경원적수량,기궤제가능시이식물-숙주접촉면적적증대사득이식물경강지폭로우숙주배경유도인소지하산생경소적궤계손상.
Objective To compare the number of DARPP-32 positive cells yield in the grafts based on the application of single cell suspension by neural micro-transplantation technique and by traditional cell delivery technique and to explore effects and mechanisms of different approaches.Methods Cells derived from the whole ganglionic eminence of E15 rat embryos,ubiquitously expressing Green Fluorescent Protein(GFP) were implanted into unilaterally QA-lesioned rat striatum in a single-tract with an ultra-thin glass capillary with an outer diameter of 50 μm or using traditional cannula tip with a diameter of 500 μm.Results Histological assessment at 4 months after transplantation showed that there was about two-fold DARRP-32 positive striatal-like neurons in the micro-transplantation group(TT group) than that in the traditional group(MT group).Total graft volume was similar in both groups[(2.8±0.2) mm3 vs(2.5±0.4)mm3,F =0.25,P ＞ 0.05].And DARRP-32positive plaque volume[(0.6±0.1) mm3 vs(0.6±0.2) mm3,F =0.90,P ＞ 0.05]and TH staining plaque volume[(1.0±0.1) mm3 vs(0.7±0.1)mm3,F =1.44,P ＞ 0.05]also had the same performance in both groups.Number of DARRP-32 positive cells was calculated by Abercrombie correction formula,and the result showed that the number of DARRP-32 positive cells in MT group was two-fold of that in TT group[(20.1 ×103±1.2 × 103) vs(9.8 × 103±3.2 × 103),F =8.62,P ＜ 0.05].Higher DARRP-32 positive cells in MT group indicated that grafts had a better condition of growth and development.Conclusion Micro-transplantation approach can increase the number of new born striatal-like neurons,potentially due to the enlargement of the graft-host border area intensifying the graft's exposure to host derived factors and the minimized mechanical injury.
