摘要
取生后２～５ｄ的ＳＤ大鼠四肢骨骼肌制备细胞悬液，并以１×１０６细胞密度放入３５ｍｍ培养皿中培养５～７ｄ，使其发育为成肌细胞及肌管细胞。按２：３比例将两种细胞混合，以大约１０６细胞数分两点（总量为１５μｌ）移植到４２只成年大鼠纹状体中。有４例动物移植前用荧光染料ＤｉＩ对肌细胞染色。移植后动物分别存活２周、４周、８周、１６周、６个月和１２个月。根据不同存活期将大鼠分组灌注取脑、冠状切片。切片经横纹肌动蛋白和肌红蛋白免疫组化染色，移植部均呈阳性反应。所有动物的移植物在肉眼及镜下均显示存活良好，除有部分胶质细胞浸润外并未发现移植物有明显缩小、扩大及肿瘤形成。另将移植后２个月及４个月共４只大鼠脑内移植物取出进行再培养，发现仍有部分肌细胞存活。本研究提示骨骼肌细胞经体外培养后移植到脑内可长期存活，并因此可作为基因治疗某些中枢神经系统疾病的一种运载工具。
The stem cells of skeletal muscle from SD rats in an age of 2-5 days were grown in a 35-mm plate with complete medium for 5- 7 days and myoblasts and myotubes were harvested.These primary muscle cells were implanted stereotaxically into the caudate nucleus of 42 adult SD rats,according to a ratio of cell number that was 2 : 3 for myoblasts and myotubes. The implanted amount was 15μl containing 106 cells for each case. The grafts were checked after different postoperative survivals from 2 weeks to 12 months, by using different methods including immnuocytochemistry. Actin and myosin staining confirmed that brains grafted with cell cultures contained actin-and myosin-positive muscle cells well survived even after 6 months. Under the light microscopy all grafts showed no phenomena of dystrophy or tumour formation except for some gliosis.The grafts with 2 and 4 months of survival in 4 cases were taken out from the hosts and recultured in vitro.It was found that those recultured muscle cells grew well and showed to be myosin-positive. The results of present study indicated that the primary muscle cells had long-term survival in rat brain and this technique would be useful for studies on gene therapy for some degeneration diseases in the brain.
出处
《解剖学报》
CAS
CSCD
北大核心
1996年第1期6-9,共4页
Acta Anatomica Sinica
基金
国家自然科学基金
北京市自然科学基金
北京市科委基金
关键词
肌细胞培养
脑移植
Primary muscle cells
Intracerebral grafting
Rat
