解剖学报
解剖學報
해부학보
ACTA ANATOMICA SINICA
2010年
1期
165-168
,共4页
精原细胞%改良显微注射器%睾丸输出管%移植%小鼠
精原細胞%改良顯微註射器%睪汍輸齣管%移植%小鼠
정원세포%개량현미주사기%고환수출관%이식%소서
Spermatogonial cells%Modified microinjection pipette%Testicular efferent duct%Transplantation%Mouse
目的 对现有小鼠精原细胞移植技术进行改进,建立更具操作性,易于推广的新方法.方法 分离含绿色荧光蛋白(GFP)的rC57BL6/tg14 (act-EGFO-Osb Y01)小鼠精原细胞作为供体细胞,应用自制的显微注射器将其从睾丸的精子输出管处,以锥虫蓝作为指示剂直接注入野生型预处理C57BL6小鼠精细管内完成移植.应用荧光显微镜观察移植效果并进行组织学分析.结果 移植后受体小鼠睾丸出现绿色荧光信号,组织学检查发现精细管中荧光信号显著高于周围组织,移植的供体小鼠(GFP)的睾丸细胞在受体小鼠睾丸中形成克隆和形成精子,而预处理的野生型小鼠睾丸精细管未发现.结论 含GFP蛋白的rC57BL6/tg14小鼠精原细胞在受体小鼠体内移植成功,此改进方法在简化精原细胞移植技术的同时可保证移植质量.
目的 對現有小鼠精原細胞移植技術進行改進,建立更具操作性,易于推廣的新方法.方法 分離含綠色熒光蛋白(GFP)的rC57BL6/tg14 (act-EGFO-Osb Y01)小鼠精原細胞作為供體細胞,應用自製的顯微註射器將其從睪汍的精子輸齣管處,以錐蟲藍作為指示劑直接註入野生型預處理C57BL6小鼠精細管內完成移植.應用熒光顯微鏡觀察移植效果併進行組織學分析.結果 移植後受體小鼠睪汍齣現綠色熒光信號,組織學檢查髮現精細管中熒光信號顯著高于週圍組織,移植的供體小鼠(GFP)的睪汍細胞在受體小鼠睪汍中形成剋隆和形成精子,而預處理的野生型小鼠睪汍精細管未髮現.結論 含GFP蛋白的rC57BL6/tg14小鼠精原細胞在受體小鼠體內移植成功,此改進方法在簡化精原細胞移植技術的同時可保證移植質量.
목적 대현유소서정원세포이식기술진행개진,건립경구조작성,역우추엄적신방법.방법 분리함록색형광단백(GFP)적rC57BL6/tg14 (act-EGFO-Osb Y01)소서정원세포작위공체세포,응용자제적현미주사기장기종고환적정자수출관처,이추충람작위지시제직접주입야생형예처리C57BL6소서정세관내완성이식.응용형광현미경관찰이식효과병진행조직학분석.결과 이식후수체소서고환출현록색형광신호,조직학검사발현정세관중형광신호현저고우주위조직,이식적공체소서(GFP)적고환세포재수체소서고환중형성극륭화형성정자,이예처리적야생형소서고환정세관미발현.결론 함GFP단백적rC57BL6/tg14소서정원세포재수체소서체내이식성공,차개진방법재간화정원세포이식기술적동시가보증이식질량.
Objective To improve the current method of spermatogonial cells transplantation, and make the new method become more operative and easy to carry out. Methods The spermatogonial cells in rC57BL6/tg14 (act-EGFO-Osb Y01) mice that expresse GFP protein were collected as the donor cells, and by using a modified syringe, and then were injected into the seminiferous tubules of pretreated wild type GFP-null mice through testicular efferent duct. The transplantation outcome was evaluated by trypan blue stainting and fluorescent microscopic examination and pathohistological analysis of transplanted testis. Results The transplanted testis of recipient mice showed green fluorescence signal, and the signal of seminiferous tubules was found significantly higher than that of the surrounding tissues. The transplanted GFP-positive cells generated colonies and spermatogenesis but pretreated wild type GFP-null mice were not found. Conclusion The expressed GFP protein spermatogonial cells in rC57BL6/tg14 mice were successfully transplanted in the wild type GFP-null recipient mice, fourthermore the improved transplantation method simplified the triditional one and achieved the same transplantation results.
