人骨髓来源细胞外基质对人牙周膜干细胞增殖的影响

Effects of human bone marrow cells-derived extracellular matrix on the proliferation of human periodontal ligament stem cells

背景:研究发现人来源的骨髓细胞外基质能促进人牙周膜干细胞增殖并保持干细胞的特性。目的:初步观察人骨髓来源的细胞外基质对人牙周膜干细胞增殖能力影响。方法:分离人正常牙周膜细胞及颌骨骨髓细胞,制备骨髓细胞外基质膜片。采用有限稀释法克隆培养纯化人牙周膜干细胞,透射电镜观察细胞超微结构。取P2代人牙周膜干细胞与骨髓细胞来源细胞外基质共同培养,以普通培养基对照,采用CCK8法及细胞周期流式技术检测人骨髓细胞来源细胞外基质对人牙周膜干细胞增殖能力的影响。结果与结论:实验组人牙周膜干细胞较对照组相比细胞增殖能力显著增强(P<0.05),且细胞符合其生物学形态及生物学特性,生长状态良好。说明实验建立起的骨髓细胞外基质能够促进牙周膜干细胞的增殖,是一种扩增干细胞的有效方法。

BACKGROUND：Human bone marrow cells-derived extracellular matrix can promote proliferation of human periodontal ligament stem cells and maintain stem cellproperties. OBJECTIVE：To preliminarily investigate the effect of human bone marrow cells-derived extracellular matrix on the proliferation of human periodontal ligament stem cells. METHODS：Human periodontal ligament stem cells and bone marrow cells were separately derived from human periodontal tissue and jaw bone marrow, and human bone marrow cells-derived extracellular matrix was prepared. Human periodontal ligament stem cells were cultured and purified using limited dilution cloning method, and transmission electron microscope was used for ultrastructure observation. Human periodontal ligament stem cells at passage 2 were cultured with human bone marrow cells-derived extracellular matrix and normal culture medium （control group）. The cellcounting kit-8 and flow cytometry were used to determine the proliferation potential of human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix.RESULTS AND CONCLUSION：Compared with the control group, human periodontal ligament stem cells cultured on human bone marrow cells-derived extracellular matrix had a superior capacity of proliferation （P〈0.05）, and the cells met their morphological and biological characteristics, and grew in good conditions. Human bone marrow cells-derived extracellular matrix is a promising matrix for large-scale expanding human periodontal ligament stem cells for future use in stem cel-based therapy.