摘要
背景:细胞种植密度是影响干细胞分化的因素之一,对于细胞种植密度在骨髓间充质干细胞向神经元样细胞分化过程中的作用尚缺乏深入研究。目的:观察细胞种植密度对骨髓间充质干细胞诱导向神经元样细胞分化的影响。方法:采用贴壁培养法分离大鼠骨髓间充质干细胞,传至第4代后将其按2×102,2×103,4×103,8×103,2×104,4×104/cm2种植于六孔板,每组均加入碱性成纤维细胞生长因子+表皮生长因子+维甲酸诱导向神经元样细胞分化,并通过免疫组织化学染色鉴定,计算每组细胞出现神经元样细胞的比例,比较各组的分化率。结果与结论:各组骨髓间充质干细胞加入诱导剂后均出现神经元样细胞,Nestin、NSE、GFAP细胞化学染色呈阳性。不同种植密度组出现神经元样细胞比例不同,以8×103/cm2组神经元样细胞比例最高,且神经元样存活时间最长,达7d。结果说明骨髓间充质干细胞向神经元样细胞分化与细胞接种密度有关,过高或过低细胞密度均不利分化。
BACKGROUND：Cell plant density is one of the factors that affect stem cell differentiation. To date,there is still lack of further study about the effect of cell plant density on bone marrow mesenchymal stem cells differentiation into neuronal-like cells. OBJECTIVE：To investigate the effect of cell density on the differentiation of rat bone marrow mesenchymal stem cells into neuronal-like cells. METHODS：Rat bone marrow mesenchymal stem cells were isolated by use of adherent culture method. Following four passages,cells were cultured in six-well plates at a density of 2×10^2/cm^2,2×103/cm^2,and 4×10^3/cm^2,8×10^3/cm^2,2×10^4/cm^2,4×10^4/cm^2. Cells in each group were differentiated into neuronal-like cells under the induction of basic fibroblast growth factor ＋ epidermal growth factor ＋ RA,and were identified by immunohistochemical staining method. The proportion of neuronal-like cells in each group was calculated,and the differentiation rate was compared. RESULTS AND CONCLUSION：Neuronal-like cells were observed after adding inducer in bone marrow mesenchymal stem cells,and Nestin,neuron-specific enolase,glial fibrillary acidic protein staining was positive. Different cell plant densities induced different ratios of neuron-like cells. 8×10^3/cm^2 plant density yielded highest proportion of neuronal-like cells and can survive as long as 7 days. The results demonstrated that the differentiation of bone mesenchymal stem cells into neuron-like cells was correlated with cell plant density. Too high or too low cell plant density was detrimental to differentiation.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2010年第32期5908-5912,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
