摘要
背景:细胞外基质可以在体外模拟细胞微环境,使干细胞在体外大量增殖的同时具有良好的干细胞特性。目的:制备人骨髓细胞外基质并分析其结构成分。方法:取第4代人骨髓细胞培养14 d,在最后8 d换成膜诱导液,经脱细胞处理后制备人骨髓细胞外基质。倒置显微镜和扫描电镜观察人骨髓细胞外基质表面形态。免疫荧光染色观察脱细胞处理前后Ⅰ型胶原和二聚糖的变化。将人牙周膜干细胞接种于人骨髓细胞外基质、纤粘连蛋白包被的六孔板和普通培养板,比较不同基质对人牙周膜干细胞细胞形态和黏附的影响。结果与结论:化学物理联合脱细胞处理可以获得结构完整的人骨髓细胞外基质膜片,Ⅰ型胶原和二聚糖在去细胞处理前后结构和含量无明显差异。接种在细胞外基质上生长的人牙周膜干细胞,按照细胞外基质的轨道有序生长,不同于原有细胞形态;相同时间人牙周膜干细胞在细胞外基质上贴壁数量较多。表明有效的脱细胞处理可以得到结构完整的细胞外基质网状支架,相关蛋白成分未因脱细胞而明显丧失,细胞外基质影响接种细胞的形态、促进细胞黏附。提示可利用细胞外基质模型模拟体内干细胞生长微环境,在体外获得大量高质量的成体干细胞。
BACKGROUND：Extracellular matrix can simulate microenvironment and make the stem cells proliferate maintaining the characteristics of stem cells wel in vitro. OBJECTIVE：To prepare the extracellular matrix from human bone marrow cells and to analyze its microstructure and composition preliminarily. METHODS：Human bone marrow cells of passage 4 were cultured for 14 days, and the induction medium was used during the last 8 days. After decellularization, cells were removed to prepare human bone marrow cells-derived extracellular matrix. The surface morphology of human bone marrow cells-derived extracellular matrix was observed by inverted microscope and scanning electron microscope. Changes of col agen I and biglycan before and after decellularization were observed by immunofluorescence staining. Human periodontal ligament stem cells were seeded onto human bone marrow cells-derived extracellular matrix, fibronectin coated 6-wel plate and normal culture plate to compare the influence of different matrix on cellmorphology and adhesion. RESULTS AND CONCLUSION：We obtained intact human bone marrow cells-derived extracellular matrix by chemical combined with physical decellularization. The structure and amount of col agen I and biglycan had not been compromised dramatical y after decellularization. Human periodontal ligament stem cells growing on the human bone marrow cells-derived extracellular matrix developed in accordance with the orbit of the extracellular＆amp;nbsp;matrix, differing from the original cellmorphology. There were more human periodontal ligament stem cells adhering to the extracellular matrix during the same time. These findings indicate that effective decellularization can produce intact the extracellular matrix membrane without destroying its microstructure. Extracellular matrix protein is not compromised due to decellularization. The extracellular matrix affects cellmorphology and promotes celladhesion. We can use the extracellular matrix model to simulate stem cellmicroenvironment and thereafter, acquire a large number of adult stem cells with high quality in vitro.
出处
《中国组织工程研究》
CAS
CSCD
2014年第10期1629-1634,共6页
Chinese Journal of Tissue Engineering Research
基金
2011年国家自然科学基金青年科学基金项目(81100750)~~
