Mao Xi, Chu Chen-Ling, Mao Zhao, Wang Jian-Jun
Department of Biological Science and Technology and State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 22 Hankou Road, Nanjing 210093, China.
Tissue Cell. 2005 Oct;37(5):349-57. doi: 10.1016/j.tice.2005.05.003.
The purposes of this study were to develop a new cultural method for the rat bone marrow stromal cells (MSCs) to differentiate into osteoblasts well in vitro, and to investigate the feasibility of using MSCs as seed cells and three-dimensional porous nano-hydroxylapatite as scaffolds for constructing tissue-engineered bone. MSCs of rats were isolated, cultured, induced to differentiate into osteoblasts, and then observed with inverted microscopy. Histochemical staining and radio-immunological analysis were applied for identifying MSCs. Whereafter MSCs were seeded onto three-dimensional porous nano-hydroxylapatite scaffolds, and scanning electron microscopy was applied to evaluate their growth on scaffolds. Results showed that MSCs were typical fibroblast-like and possessed a better proliferating capability; the activity of alkaline phosphatase (ALP) and the secretion of osteocalcin of MSCs were produced gradually and increased continuously; the cells seeded on three-dimensional porous nano-hydroxylapatite scaffolds adhered, proliferated and differentiated well. These results demonstrated that the new improved culture method had the advantages of short isolating time, less risk of contamination and higher efficiency and accordingly was conducive to MSCs proliferating and differentiating into osteoblasts, and that it was advantageous to constructing tissue-engineered bone using MSCs as seed cells and three-dimensional porous nano-hydroxylapatite as scaffolds.
本研究的目的是开发一种新的培养方法，使大鼠骨髓基质细胞（MSCs）在体外能良好地分化为成骨细胞，并研究将MSCs作为种子细胞以及使用三维多孔纳米羟基磷灰石作为支架构建组织工程骨的可行性。分离、培养大鼠的MSCs，诱导其分化为成骨细胞，然后用倒置显微镜观察。采用组织化学染色和放射免疫分析来鉴定MSCs。之后将MSCs接种到三维多孔纳米羟基磷灰石支架上，并用扫描电子显微镜评估其在支架上的生长情况。结果显示，MSCs呈典型的成纤维细胞样，具有较好的增殖能力；MSCs的碱性磷酸酶（ALP）活性和骨钙素分泌逐渐产生并持续增加；接种在三维多孔纳米羟基磷灰石支架上的细胞黏附、增殖和分化良好。这些结果表明，新改进的培养方法具有分离时间短、污染风险小和效率高的优点，因此有利于MSCs增殖并分化为成骨细胞，且有利于以MSCs作为种子细胞、三维多孔纳米羟基磷灰石作为支架构建组织工程骨。
