Department of Biomaterials, Field of Tissue Engineering, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawara-cho Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
J Biomater Sci Polym Ed. 2010;21(5):609-21. doi: 10.1163/156856209X434638.
The objective of this study was to evaluate the proliferation and chondrogenic differentiation of rat bone marrow-derived mesenchymal stem cells (MSCs) cultured with gelatin hydrogel microspheres of cell scaffold which can release transforming growth factor-beta1 (TGF-beta1). Gelatin was dehydrothermally cross-linked in different conditions in a water-in-oil emulsion state to obtain gelatin hydrogel microspheres with different water content. The microspheres functioned not only as the scaffold of MSC, but also the carrier matrix of TGF-beta1 release. The MSC proliferation depended on the water content of microspheres. Higher MSC proliferation was observed for the gelatin microspheres with lower water content. When cultured with the gelatin hydrogel microspheres, MSC formed their aggregates, in contrast to culturing with hydrogel sheets. The cell viability was significantly high compared with that of the hydrogel sheet. The production of sulfated glycosaminaglycan (sGAG) from MSC was examined as a measure of chondrogenic differentiation, after their culturing in a normal and chondrogenic differentiation media. For both the cultures, the amount of sGAG produced was significantly higher for MSC cultured with the gelatin microspheres than that of the gelatin sheet. Stronger differentiation of MSC was achieved in culture with the microspheres incorporating TGF-beta1 than that of MSC cultured in the medium containing the same amount of TGF-beta1. It is concluded that the gelatin hydrogel microspheres function well as both the scaffold of MSC and the matrix of TGF-beta1 release, resulting in enhanced MSC aggregation and the consequent promotion of cell proliferation and differentiation.
本研究的目的是评估大鼠骨髓间充质干细胞（MSCs）在可释放转化生长因子-β1（TGF-β1）的细胞支架明胶水凝胶微球中培养时的增殖和软骨分化情况。明胶在油包水乳液状态下以不同条件进行热交联，以获得具有不同含水量的明胶水凝胶微球。这些微球不仅可以作为 MSC 的支架，还可以作为 TGF-β1 释放的载体基质。MSC 的增殖取决于微球的含水量。具有较低含水量的明胶微球观察到更高的 MSC 增殖。当与明胶水凝胶微球共培养时，MSC 形成其聚集物，与水凝胶片的培养相反。与水凝胶片相比，细胞活力明显更高。作为软骨分化的衡量标准，检测了 MSC 在正常和软骨分化培养基中培养时硫酸化糖胺聚糖（sGAG）的产生。对于两种培养物，与明胶片相比，用明胶微球培养的 MSC 产生的 sGAG 量明显更高。在包含 TGF-β1 的微球中培养的 MSC 的分化强于在含有相同量 TGF-β1 的培养基中培养的 MSC 的分化。结论是，明胶水凝胶微球可以很好地充当 MSC 的支架和 TGF-β1 释放的基质，从而增强 MSC 的聚集，并促进细胞增殖和分化。
