摘要
背景:肌营养不良症是一种渐进性致死性X连锁隐性遗传性肌肉疾病,目前无特效治疗。肌营养不良症模型鼠(mdx小鼠)的骨髓基质细胞增殖及定向分化能力是否正常,自身骨髓移植是否合适还有待研究。目的:观察mdx小鼠骨髓基质细胞体外培养时的增殖及多向分化能力。方法:取mdx小鼠与C57小鼠胫股骨骨髓基质细胞体外培养,经吉姆萨染色后观察其形成成纤维细胞集落形成单位的能力;通过不同诱导液使骨髓基质细胞定向分化为成骨、成脂、成肌细胞,观察其形态学特性;并分别用Vonkossa染色、油红O染色、免疫荧光检测desmin阳性细胞对已分化细胞进行鉴定和分化率比较;培养1周时,提取分化细胞总RNA,反转录后,用real-time PCR检测各分化细胞相关基因表达。结果与结论:mdx小鼠骨髓基质细胞形成的成纤维细胞集落形成单位数目和体积均小于C57小鼠。其成骨、成肌分化的效率均明显低于C57小鼠(P<0.01),两种小鼠的骨髓基质细胞成脂分化效率差异无显著性(P>0.05)。real-time PCR检测结果显示,与C57小鼠相比,mdx小鼠的骨髓基质细胞成骨、成肌基因表达均有不同程度下降,而成脂基因表达无明显差异。结果提示,mdx小鼠的骨髓基质细胞体外培养时的增殖及定向分化能力较C57小鼠下降,与Dystrophin基因缺失有关,mdx小鼠自体骨髓移植将会受限。
BACKGROUND：Duchenne＇s muscular dystrophy （DMD） is a fatal recessive X-linked form of muscular dystrophy characterized by progressive muscular degeneration with no certain treatment.However, the proliferation and multipotent differentiation potential of bone marrow stromal cells （BMSCs） from DMD model mice （mdx mice） and the effectiveness of BMSCs self-transplantation are needed to be further studied.OBJECTIVE： To investigate the proliferation and multipotent differentiation potential of BMSCs from mdx mice during in vitro culture.METHODS： Both C57 and mdx mice were killed to obtain the BMSCs for culturing in vitro.After Giemsa＇s staining, the colony forming unit-fibroblast （CFU-f） of BMSCs assay was performed.With specific inductive mediums, we succeeded to induce the BMSCs to differentiate into osteogenesis, adipogenesis, myogenesis respectively.Their morphological characteristics were observed with microscope.Von kossa staining, oil red O staining and immunofluorescence for desmin were utilized to identify the differentiated BMSCs respectively and quantify their differentiation efficiency.After the BMSCs were induced for 1 week, the cellular total RNA of differentiated BMSCs was extracted, and then reverse transcription was performed.Real-time polymerase chain reaction （PCR） was used to quantify the gene expression of differentiated BMSCs.RESULTS AND CONCLUSION： Compared with C57 mice, both number and volume of CFU-f of BMSCs from mdx mice were smaller.Compared to C57 mice, the efficiency of osteogenesis and myogenesis of BMSCs from mdx mice was significantly lower （P〈0.01）.However, the efficiency of adipogenesis of BMSCs from both groups had no statistic difference （P〉0.05）.Real-time PCR showed that both of osteogenic and myogenic gene expression of BMSCs from mdx mice decreased respectively compared to C57 mice.However, adipogenic gene expression from two groups had no difference.The results indicated that proliferation and multipotent differentiation potentials of BMSCs from mdx mice declined compared with C57 mice.The defection of Dystrophin gene may contribute to it.Therefore, the autoplastic BMSCs transplantation of mdx mice will be interfered.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第36期6651-6656,共6页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然基金资助:GILZ对骨髓间质干细胞分化的调控及其机制的研究(30870852)
GILZ转导的MSC移植治疗DMD模型的研究(30971026)~~
