Differentiation of mesenchymal stem cells into osteoblasts on honeycomb collagen scaffolds
Differentiation of mesenchymal stem cells into osteoblasts on honeycomb collagen scaffolds
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DOI：
10.1002/bit.20939
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发表时间：
2006-10-20
影响因子：
3.8
通讯作者：
Miyata, Teruo
中科院分区：
文献类型：
--
作者：
George, Joseph;Kuboki, Yoshinori;Miyata, Teruo
使用活细胞的组织工程正在成为组织或器官移植的替代方案。成体间充质干细胞在特定的生长因子作用下，可分化为脂肪细胞、软骨细胞或成骨细胞等多系细胞。在本研究中，我们研究了蜂窝胶原支架对骨髓间充质干细胞向成骨细胞的黏附、分化和增殖的影响。从6周龄大鼠股骨骨髓中分离出间充质干细胞，在不含β-甘油磷酸和地塞米松的α-MEM培养液中培养。以牛真皮基质为原料制备蜂窝状胶原盘，紫外光辐射交联，加热灭菌。在干细胞接种之前，蜂窝圆盘被放在培养皿上。细胞迅速附着在蜂窝状胶原支架上，分化并增殖为成骨细胞。对分化后的成骨细胞进行形态观察和碱性磷酸酶活性测定。成骨细胞还在培养中合成了缺钙羟基磷灰石(伪羟基磷灰石)晶体。用Von Kossa染色确定了矿化，用X射线衍射仪对晶体进行了分析。光镜和DNA测量显示，分化的成骨细胞在蜂窝状胶原支架上增殖成几层。结果表明，蜂窝胶原海绵是骨髓间充质干细胞向成骨细胞分化和增殖的良好支架材料。这些数据进一步证明，蜂窝状胶原是一种有效的组织工程应用底物，在干细胞技术和细胞治疗的前沿领域非常有用。(C)2006年威利期刊公司。
Tissue engineering using living cells is emerging as an alternative to tissue or organ transplantation. The adult mesenchymal stem cells can be differentiated into multilineage cells, such as adipocytes, chondrocytes, or osteoblasts when cultured with specific growth factors. In the present investigation, we have studied the effect of honeycomb collagen scaffolds for the adhesion, differentiation and proliferation of bone marrow-derived mesenchymal stem cells into osteoblasts. Mesenchymal stem cells were isolated from 6-week old albino rat femur bone marrow, and cultured in alpha-MEM medium without beta-glycerophosphate and dexamethasone. Honeycomb collagen discs were prepared from bovine dermal atelocollagen, cross-linked by UV-irradiation and sterilized by heat. The honeycomb discs were placed on the culture dishes before seeding the stem cells. The cells attached quickly to the honeycomb collagen scaffold, differentiated and proliferated into osteoblasts. The differentiated osteoblasts were characterized by morphological examination and alkaline phosphatase activity. The osteoblasts also synthesized calcium-deficient hydroxyapatite (pseudo-hydroxyapatite) crystals in the culture. The mineralization was confirmed by Von Kossa staining and the crystals were analyzed by X-ray diffraction. Light microscopy and DNA measurements showed that the differentiated osteoblasts multiplied into several layers on the honeycomb collagen scaffold. The results demonstrated that the honeycomb collagen sponge is an excellent scaffold for the differentiation and proliferation of mesenchymal stem cells into osteoblasts. The data further proved that honeycomb collagen is an effective substrate for tissue engineering applications, and is very useful in the advancing field of stem cell technology and cell-based therapy. (c) 2006 Wiley Periodicals, Inc.