摘要
目的寻求大鼠肌源细胞体外大量培养及鉴定的方法,为临床直接使用自体肌源细胞注射治疗肌组织损伤疾病奠定实验基础。方法利用胶原酶和胰蛋白酶消化3周龄大鼠腓肠肌,应用选择性培养基抑制非肌源细胞生长,采取改良的差速贴壁法纯化肌源细胞,通过形态观察、免疫组化及RT-PCR进行鉴定。结果使用F-10培养基获得了大量高纯度的大鼠肌源细胞;倒置显微镜下可观察到聚集成团和散在分布的两种形态的肌源细胞;免疫组化显示肌源细胞Desm in阳性率约90%,聚集成团的为CD34+,大部分单个存在的为CD34-;RT-PCR显示传代细胞中均有Desm in基因的表达。结论此培养方法可在体外获得大量大鼠肌源细胞,有助于基因工程及组织工程等领域的研究。
Objective To explore an efficient primary culture method for rat muscle-derived cells（MDC） so as to provide experimental basis for treating muscle tissue injury disease by injecting MDC.Methods MDC were isolated from gastrocnemius muscle biopsies of 3-week-old Sprague-Dawlay（SD） rats and digested by collagenase and trypsin,which was then purified by different adhesion time and selective media.The cells were identified by morphology,immunohistochemistry and RT-PCR.Results A large number of high-purified MDC were enriched by using F-10 culture media.Aggregated cells and dispersed cells were observed by inverted microscope.The positive rates of Desmin were about 90% in MDC.Immunohistochemisty showed that CD34＋ were expressed in the aggregated cells and CD34-were expressed in most of the dispersed cells.RT-PCR confirmed the expression of Desmin gene in passage cells.Conclusion It＇s feasible to culture MDC primarily in vitro and these cells can be used in genetic and tissue engineering field.
出处
《同济大学学报（医学版）》
CAS
2009年第2期19-23,共5页
Journal of Tongji University(Medical Science)
基金
国家自然科学基金资助项目(30571866)
关键词
肌源细胞
培养
纯化
大鼠
muscle-derived cells
culture
purification
rats
