Preparation of stem cell aggregates with gelatin microspheres to enhance biological functions
Preparation of stem cell aggregates with gelatin microspheres to enhance biological functions
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DOI：
10.1016/j.actbio.2011.04.013
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发表时间：
2011-07-01
影响因子：
9.7
通讯作者：
Tabata, Yasuhiko
中科院分区：
文献类型：
--
作者：
Hayashi, Kentaro;Tabata, Yasuhiko
本研究的目的是通过使用明胶水凝胶微球提高培养的大鼠骨髓间充质干细胞（MSC）的活力和成骨分化。明胶在油包水乳液状态下在140 ℃下脱氢热交联48小时。当与明胶水凝胶微球一起培养时，包被有聚（乙烯醇）MSC的96孔板的圆形、U形底威尔斯孔形成均匀结合微球的聚集体。与没有微球形成的聚集体相比，细胞聚集体的活力显著更高。MSC在聚集体中的增殖依赖于掺入的微球的数量和直径。对于掺入更多数量的较大明胶微球的聚集体，观察到更高的MSC增殖。当评价作为有氧糖酵解的量度时，MSC中L-乳酸产生/葡萄糖消耗的比率对于与明胶微球一起培养的MSC显著低于那些没有微球的MSC。MSC碱性磷酸酶（ALP）和硫酸化糖胺聚糖（sGAG）的生产进行了检查，以评估其潜在的成骨和软骨分化。与无微球培养的MSC相比，用明胶微球培养的MSC聚集体产生的ALP的量显著更高。另一方面，对于含有微球的MSC聚集体，产生的sGAG的量显著较低。可以得出结论，明胶水凝胶微球的掺入防止聚集的MSC遭受缺氧，导致增强的MSC聚集和细胞增殖和成骨分化。(C)2011 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
The objective of this study is to improve the viability and osteogenic differentiation of cultured rat bone marrow-derived mesenchymal stem cells (MSC) by the use of gelatin hydrogel microspheres. Gelatin was dehydrothermally crosslinked at 140 degrees C for 48 h in a water in oil emulsion state. When cultured with the gelatin hydrogel microspheres in round, U-bottomed wells of 96-well plates coated with poly(vinyl alcohol) MSC formed aggregates homogeneously incorporating the microspheres. The viability of the cell aggregates was significantly higher compared with that of aggregates formed without microspheres. MSC proliferation in the aggregates depended on the number and diameter of the incorporated microspheres. Higher MSC proliferation was observed for aggregates incorporating a greater number of larger gelatin microspheres. When evaluated as a measure of aerobic glycolysis the ratio of L-lactic acid production/glucose consumption in MSC was significantly lower for MSC cultured with gelatin microspheres than those without microspheres. MSC production of alkaline phosphatase (ALP) and sulfated glycosaminaglycan (sGAG) was examined to evaluate their potential osteogenic and chondrogenic differentiation. The amount of ALP produced was significantly higher for MSC aggregates cultured with gelatin microspheres than that of MSC cultured without microspheres. On the other hand, the amount of sGAG produced was significantly lower for MSC aggregates containing microspheres. It is concluded that the incorporation of gelatin hydrogel microspheres prevents the aggregated MSC suffering from a lack of oxygen, resulting in enhanced MSC aggregation and cell proliferation and osteogenic differentiation. (C) 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.