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目的观察壳聚糖薄膜培养间充质干细胞(MSCs)成为球状体的可能性及其对MSCs干性标记基因的影响。方法在壳聚糖薄膜培养MSCs,用MTT法研究其细胞的存活率和细胞增殖;用实时定量PCR检测MSCs的干性基因的表达,研究壳聚糖培养的MSCs分化成神经细胞和软骨形成的可能性。结果壳聚糖薄膜培养的MSCs能形成球状体。壳聚糖培养比较二维培养(2D)的MSCs的干性基因Oct-4、SOX和Nanog等明显增加。壳聚糖薄膜上培养的MSC细胞更具有分化成神经细胞和软骨细胞的潜能。结论壳聚糖薄膜培养的MSCs能够保持其干性,适合用于包括脑栓塞和软骨组织疾病的细胞治疗。
OBJECTIVE: To investigate the possibility of chitosan membrane-derived mesenchymal stem cells (MSCs) becoming spheroids and their effect on the expression of MSCs dry marker genes. Methods MSCs were cultured on chitosan film, cell viability and cell proliferation were studied by MTT method. The expression of dry genes in MSCs was detected by real-time quantitative PCR, and the differentiation of MSCs cultured in chitosan into nerve cells and cartilage possibility. Results Chitosan cultured MSCs could form spheroids. Chitosan culture significantly increased the expression of Oct-4, SOX and Nanog in the two-dimensional culture MSCs. MSC cells cultured on chitosan film have more potential to differentiate into nerve cells and chondrocytes. Conclusion Chitosan cultured MSCs can maintain their dryness and are suitable for cell therapy including cerebral embolism and cartilage diseases.